Compounds as inhibitors of hepatitis C virus NS3 serine protease

ABSTRACT

The present invention discloses novel compounds which have HCV protease inhibitory activity as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising such compounds as well as methods of using them to treat disorders associated with the HCV protease.

This application is a divisional of U.S. application Ser. No.11/065,509, filed Feb. 24, 2005, and claims the benefit of U.S.Provisional Application Ser. No. 60/548,507 filed Feb. 27, 2004.

FIELD OF THE INVENTION Background of the Invention

Hepatitis C virus (HCV) is a (+)-sense single-stranded RNA virus thathas been implicated as the major causative agent in non-A, non-Bhepatitis (NANBH), particularly in blood-associated NANBH (BB-NANBH)(see, International Patent Application Publication No. WO 89/04669 andEuropean Patent Application Publication No. EP 381 216). NANBH is to bedistinguished from other types of viral-induced liver disease, such ashepatitis A virus (HAV), hepatitis B virus (HBV), delta hepatitis virus(HDV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), as well asfrom other forms of liver disease such as alcoholism and primary biliarcirrhosis.

Recently, an HCV protease necessary for polypeptide processing and viralreplication has been identified, cloned and expressed. (See, e.g. U.S.Pat. No. 5,712,145). This approximately 3000 amino acid polyproteincontains, from the amino terminus to the carboxy terminus, anucleocapsid protein (C), envelope proteins (E1 and E2) and severalnon-structural proteins (NS1, 2, 3, 4a, 5a and 5b). NS3 is anapproximately 68 kda protein, encoded by approximately 1893 nucleotidesof the HCV genome, and has two distinct domains: (a) a serine proteasedomain consisting of approximately 200 of the N-terminal amino acids;and (b) an RNA-dependent ATPase domain at the C-terminus of the protein.The NS3 protease is considered a member of the chymotrypsin familybecause of similarities in protein sequence, overall three-dimensionalstructure and mechanism of catalysis. Other chymotrypsin-like enzymesare elastase, factor Xa, thrombin, trypsin, plasmin, urokinase, tPA andPSA. The HCV NS3 serine protease is responsible for proteolysis of thepolypeptide (polyprotein) at the NS3/NS4a, NS4a/NS4b, NS4b/NS5a andNS5a/NS5b junctions and is thus responsible for generating four viralproteins during viral replication. This has made the HCV NS3 serineprotease an attractive target for antiviral chemotherapy. The inventivecompounds can inhibit such protease. They also can modulate theprocessing of hepatitis C virus (HCV) polypeptide.

It has been determined that the NS4a protein, an approximately 6 kdapolypeptide, is a co-factor for the serine protease activity of NS3.Autocleavage of the NS3/NS4a junction by the NS3/NS4a serine proteaseoccurs intramolecularly (i.e., cis) while the other cleavage sites areprocessed intermolecularly (i.e., trans).

Analysis of the natural cleavage sites for HCV protease revealed thepresence of cysteine at P1 and serine at P1′ and that these residues arestrictly conserved in the NS4a/NS4b, NS4b/NS5a and NS5a/NS5b junctions.The NS3/NS4a junction contains a threonine at P1 and a serine at P1′.The Cys→Thr substitution at NS3/NS4a is postulated to account for therequirement of cis rather than trans processing at this junction. See,e.g., Pizzi et al. (1994) Proc. Natl. Acad. Sci. (USA) 91:888-892,Failla et al. (1996) Folding &Design 1:35-42. The NS3/NS4a cleavage siteis also more tolerant of mutagenesis than the other sites. See, e.g.,Kollykhalov et al. (1994) J. Virol. 68:7525-7533. It has also been foundthat acidic residues in the region upstream of the cleavage site arerequired for efficient cleavage. See, e.g., Komoda et al. (1994) J.Virol. 68:7351-7357.

Inhibitors of HCV protease that have been reported include antioxidants(see, International Patent Application Publication No. WO 98/14181),certain peptides and peptide analogs (see, International PatentApplication Publication No. WO 98/17679, Landro et al. (1997) Biochem.36:9340-9348, Ingallinella et al. (1998) Biochem. 37:8906-8914,Llinàs-Brunet et al. (1998) Bioorg. Med. Chem. Lett. 8:1713-1718),inhibitors based on the 70-amino acid polypeptide eglin c (Martin et al.(1998) Biochem. 37:11459-11468, inhibitors affinity selected from humanpancreatic secretory trypsin inhibitor (hPSTI-C3) and minibodyrepertoires (MBip) (Dimasi et al. (1997) J. Virol. 71:7461-7469),cV_(H)E2 (a “camelized” variable domain antibody fragment) (Martin etal. (1997) Protein Eng. 10:607-614), and α1-antichymotrypsin (ACT)(Elzouki et al.) (1997) J. Hepat. 27:42-28). A ribozyme designed toselectively destroy hepatitis C virus RNA has recently been disclosed(see, BioWorld Today 9(217): 4 (Nov. 10, 1998)).

Reference is also made to the PCT Publications, No. WO 98/17679,published Apr. 30, 1998 (Vertex Pharmaceuticals Incorporated); WO98/22496, published May 28, 1998 (F. Hoffmann-La Roche A G); and WO99/07734, published Feb. 18, 1999 (Boehringer Ingelheim Canada Ltd.).

HCV has been implicated in cirrhosis of the liver and in induction ofhepatocellular carcinoma. The prognosis for patients suffering from HCVinfection is currently poor. HCV infection is more difficult to treatthan other forms of hepatitis due to the lack of immunity or remissionassociated with HCV infection. Current data indicates a less than 50%survival rate at four years post cirrhosis diagnosis. Patients diagnosedwith localized resectable hepatocellular carcinoma have a five-yearsurvival rate of 10-30%, whereas those with localized unresectablehepatocellular carcinoma have a five-year survival rate of less than 1%.

Reference is made to WO 00/59929 (U.S. Pat. No. 6,608,027, Assignee:Boehringer Ingelheim (Canada) Ltd.; Published Oct. 12, 2000) whichdiscloses peptide derivatives of the formula:

Reference is made to A. Marchetti et al, Synlett, S1, 1000-1002 (1999)describing the synthesis of bicylic analogs of an inhibitor of HCV NS3protease. A compound disclosed therein has the formula:

Reference is also made to W. Han et al, Bioorganic &Medicinal Chem.Lett, (2000) 10, 711-713, which describes the preparation of certainα-ketoamides, α-ketoesters and α-diketones containing allyl and ethylfunctionalities.

Reference is also made to WO 00/09558 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to WO 00/09543 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to U.S. Pat. No. 6,608,027 (Boehringer Ingelheim,Canada) which discloses NS3 protease inhibitors of the type:

wherein the various moieties are defined therein.

Current therapies for hepatitis C include interferon-α (INF _(α) ) andcombination therapy with ribavirin and interferon. See, e.g. Beremgueret al. (1998) Proc. Assoc. Am. Physicians 110(2):98-112. These therapiessuffer from a low sustained response rate and frequent side effects.See, e.g., Hoofnagle et al. (1997) N. Engl. J. Med. 336:347. Currently,no vaccine is available for HCV infection.

Reference is further made to WO 01/74768 (Assignee: VertexPharmaceuticals Inc) published Oct. 11, 2001, which discloses certaincompounds of the following general formula (R is defined therein) asNS3-serine protease inhibitors of Hepatitis C virus:

A specific compound disclosed in the afore-mentioned WO 01/74768 has thefollowing

PCT Publications WO 01/77113; WO 01/081325; WO 02/08198; WO 02/08256; WO02/08187; WO 02/08244; WO 02/48172; WO 02/08251; and pending U.S. patentapplication Ser. No. 10/052,386, filed Jan. 18, 2002, disclose varioustypes of peptides and/or other compounds as NS-3 serine proteaseinhibitors of hepatitis C virus. The disclosures of those applicationsare incorporated herein by reference thereto.

There is a need for new treatments and therapies for HCV infection.There is a need for compounds useful in the treatment or prevention oramelioration of one or more symptoms of hepatitis C.

There is a need for methods of treatment or prevention or ameliorationof one or more symptoms of hepatitis C.

There is a need for methods for modulating the activity of serineproteases, particularly the HCV NS3/NS4a serine protease, using thecompounds provided herein.

There is a need for methods of modulating the processing of the HCVpolypeptide using the compounds provided herein.

SUMMARY OF THE INVENTION

In its many embodiments, the present invention provides a novel class ofinhibitors of the HCV protease, pharmaceutical compositions containingone or more of the compounds, methods of preparing pharmaceuticalformulations comprising one or more such compounds, and methods oftreatment or prevention of HCV or amelioration of one or more of thesymptoms of hepatitis C using one or more such compounds or one or moresuch formulations. Also provided are methods of modulating theinteraction of an HCV polypeptide with HCV protease. Among the compoundsprovided herein, compounds that inhibit HCV NS3/NS4a serine proteaseactivity are preferred. The present invention discloses compounds, orenantiomers, stereoisomers, rotamers, tautomers, diastereomers andracemates of said compounds, or a pharmaceutically acceptable salt,solvate or ester of said compounds, said compounds having the having thegeneral structure shown in structural Formula 1:

wherein:

R¹ is H, OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can be the sameor different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroalkyl-,heteroaryl-, cycloalkyl-, heterocyclyl-, arylalkyl-, andheteroarylalkyl;

A and M can be the same or different, each being independently selectedfrom R, NR⁹R¹⁰, SR, SO₂R, and halo; or A and M are connected to eachother such that the moiety:

shown above in Formula I (i.e., M-L-E-A taken together) forms either athree, four, six, seven or eight-membered cycloalkyl, a four toeight-membered heterocyclyl, a six to ten-membered aryl, or a five toten-membered heteroaryl;

E is C(H) or C(R);

L is C(H), C(R), CH₂C(R), or C(R)CH₂;

R, R′, R², and R³ can be the same or different, each being independentlyselected from the group consisting of H, alkyl-, alkenyl-, alkynyl-,cycloalkyl-, heteroalkyl-, heterocyclyl-, aryl-, heteroaryl-,(cycloalkyl)alkyl-, (heterocyclyl)alkyl-, aryl-alkyl-, andheteroaryl-alkyl-; or alternately R and R′ in NRR′ are connected to eachother such that NR⁹R¹⁰ forms a four to eight-membered heterocyclyl;

Y is selected from the following moieties:

wherein Y³⁰ and Y³¹ are selected from

-   -   where u is a number 0-6;        X is selected from O, NR¹⁵, NC(O)R¹⁶, S, S(O) and SO₂;        G is NH or O; and        R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, T₁, T₂, T₃ and T₄ can be the same or        different, each being independently selected from the group        consisting of H, alkyl, heteroalkyl, alkenyl, heteroalkenyl,        alkynyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl,        arylalkyl, heteroaryl, and heteroarylalkyl, or alternately, R¹⁷        and R¹⁸ are connected to each other to form a three to        eight-membered cycloalkyl or heterocyclyl;        wherein each of said alkyl, aryl, heteroaryl, cycloalkyl or        heterocyclyl can be unsubstituted or optionally independently        substituted with one or more moieties selected from the group        consisting of: hydroxy, alkoxy, aryloxy, thio, alkylthio,        arylthio, amino, amido, alkylamino, arylamino, alkylsulfonyl,        arylsulfonyl, sulfonamido, alkyl, aryl, heteroaryl,        alkylsulfonamido, arylsulfonamido, keto, carboxy, carbalkoxy,        carboxamido, alkoxycarbonylamino, alkoxycarbonyloxy,        alkylureido, arylureido, halo, cyano, and nitro.

The above-noted statement “A and M are connected to each other such thatthe moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl” can beillustrated in a non-limiting matter as follows. Thus, for example, inthe case where A and M are connected such that the moiety:

shown above in Formula I forms a six-membered cycloalkyl(cyclohexyl),Formula I can be depicted as:

One with ordinary skill in the art will appreciate that similardepictions for Formula I can be arrived at when A and M shown above inthe moiety:

(M-L-E-A taken together) are connected to form a three, four, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl.

In the above-noted definitions of R, R′, R², and R³ preferred alkyl ismade of one to ten carbon atoms, preferred alkenyl or alkynyl is made oftwo to ten carbon atoms, preferred cycloalkyl is made of three to eightcarbon atoms, and preferred heteroalkyl, heteroaryl orheterocycloalkyl(heterocyclyl) has one to six oxygen, nitrogen, sulfur,or phosphorus atoms.

The compounds represented by Formula I, by themselves or in combinationwith one or more other suitable agents disclosed herein, can be usefulfor treating diseases such as, for example, HCV, HIV, AIDS (AcquiredImmune Deficiency Syndrome), and related disorders, as well as formodulating the activity of hepatitis C virus (HCV) protease, preventingHCV, or ameliorating one or more symptoms of hepatitis C. Suchmodulation, treatment, prevention or amelioration can be done with theinventive compounds as well as with pharmaceutical compositions orformulations comprising such compounds. Without being limited to theory,it is believed that the HCV protease may be the NS3 or NS4a protease.The inventive compounds can inhibit such protease. They can alsomodulate the processing of hepatitis C virus (HCV) polypeptide.

DETAILED DESCRIPTION

In an embodiment, the present invention discloses compounds which arerepresented by structural Formula 1 or a pharmaceutically acceptablesalt, solvate or ester thereof, wherein the various moieties are asdefined above.

In another embodiment, R¹ is NR⁹R¹⁰, and R⁹ is H, R¹⁰ is H, or R¹⁴wherein R¹⁴ is H, alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl,alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, alkenyl, alkynyl orheteroaryl-alkyl.

In another embodiment, R¹⁴ is selected from the group consisting of:

In another embodiment, R² is selected from the group consisting of thefollowing moieties:

In another embodiment, R³ is selected from the group consisting of:

wherein R³¹ is OH or O-alkyl; and

R³² is H, C(O)CH₃, C(O)OtBu or C(O)N(H)tBu.

In an additional embodiment, R³ is selected from the group consisting ofthe following moieties:

In another embodiment, G is NH.

In another embodiment, Y is selected from the following moieties:

wherein Y³² is selected from the group consisting of:

Y³⁰ and Y³¹ is selected from

-   -   wherein u is a number 0-6; and

and R¹⁹ is selected from H, alkyl, phenyl or benzyl.

In another embodiment, T₁ and T₂ can be the same or different, eachbeing independently selected from the group consisting of:

wherein T₅ and T₆ can be the same or different, each being independentlyselected from the group consisting of alkyl, aryl, cycloalkyl,heteroaryl and heterocyclyl;

or the moiety:

taken together represents

and

T₃ and T₄ can be the same or different, each being independentlyselected from:

or T₃ and T₄ taken together may form part of a four to seven memberedheterocyclic ring; in other words, the moiety T₃-N—C-T₄ may form part ofa four to seven membered heterocyclic ring.

In another embodiment, the moiety:

is selected from the following structures:

In an additional embodiment, the moiety:

is selected from the following structures:

In a still additional embodiment, the moiety:

is selected from the following structures:

In a further additional embodiment, R¹ is NHR¹⁴, where R¹⁴ is selectedfrom the group consisting of:

R² is selected from the group consisting of the following moieties:

R³ is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

and the moiety:

Yet another embodiment of the invention discloses compounds shown inTable 1, Table 2, Table 3, Table 4 and Table 5 later in thisDescription. Also shown in the Tables are the biological activities ofseveral inventive compounds (as Ki* values in nanoMolar).

In a still additional embodiment, this invention discloses the followingcompounds in Table 6:

TABLE 6 Structure Ki* (nM)

5

3.7

30

19

9

15

14

11

2.6

9

12

5.8

10

2 11399

4 11405

4 11411

5 11417

6 11401

6 11412

7 11418

7 11421

7 11395

8 11420

8 11400

9 11410

9 11402

10

As used above, and throughout this disclosure, the following terms,unless otherwise indicated, shall be understood to have the followingmeanings:

“Patient” includes both human and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight orbranched and comprising about 1 to about 20 carbon atoms in the chain.Preferred alkyl groups contain about 1 to about 12 carbon atoms in thechain. More preferred alkyl groups contain about 1 to about 6 carbonatoms in the chain. Branched means that one or more lower alkyl groupssuch as methyl, ethyl or propyl, are attached to a linear alkyl chain.“Lower alkyl” means a group having about 1 to about 6 carbon atoms inthe chain which may be straight or branched. The term “substitutedalkyl” means that the alkyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of halo, alkyl, aryl,cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, —NH(alkyl),—NH(cycloalkyl), —N(alkyl)₂, —N(alkyl)₂, carboxy and —(O)O-alkyl.Non-limiting examples of suitable alkyl groups include methyl, ethyl,n-propyl, isopropyl and t-butyl.

“Alkenyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon double bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkenyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 6 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkenyl chain. “Lower alkenyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. The term “substituted alkenyl” means that the alkenyl groupmay be substituted by one or more substituents which may be the same ordifferent, each substituent being independently selected from the groupconsisting of halo, alkyl. aryl, cycloalkyl, cyano, alkoxy and—S(alkyl). Non-limiting examples of suitable alkenyl groups includeethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyland decenyl.

“Alkynyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon triple bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkynyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 4 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkynyl chain. “Lower alkynyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. Non-limiting examples of suitable alkynyl groups includeethynyl, propynyl, 2-butynyl and 3-methylbutynyl. The term “substitutedalkynyl” means that the alkynyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of alkyl, aryl andcycloalkyl.

“Aryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 6 to about 14 carbon atoms, preferably about 6 to about10 carbon atoms. The aryl group can be optionally substituted with oneor more “ring system substituents” which may be the same or different,and are as defined herein. Non-limiting examples of suitable aryl groupsinclude phenyl and naphthyl.

“Heteroaryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 5 to about 14 ring atoms, preferably about 5 to about10 ring atoms, in which one or more of the ring atoms is an elementother than carbon, for example nitrogen, oxygen or sulfur, alone or incombination. Preferred heteroaryls contain about 5 to about 6 ringatoms. The “heteroaryl” can be optionally substituted by one or more“ring system substituents” which may be the same or different, and areas defined herein. The prefix aza, oxa or thia before the heteroarylroot name means that at least a nitrogen, oxygen or sulfur atomrespectively, is present as a ring atom. A nitrogen atom of a heteroarylcan be optionally oxidized to the corresponding N-oxide. Non-limitingexamples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl,thienyl, pyrimidinyl, pyridone (including N-substituted pyridones),isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl,pyrrolyl, pyrazolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl,pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl,imidazo[1,2-a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl,indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl,imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl,pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl,1,2,4-triazinyl, benzothiazolyl and the like. The term “heteroaryl” alsorefers to partially saturated heteroaryl moieties such as, for example,tetrahydroisoquinolyl, tetrahydroquinolyl and the like.

“Aralkyl” or “arylalkyl” means an aryl-alkyl-group in which the aryl andalkyl are as previously described. Preferred aralkyls comprise a loweralkyl group. Non-limiting examples of suitable aralkyl groups includebenzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parentmoiety is through the alkyl.

“Alkylaryl” means an alkyl-aryl-group in which the alkyl and aryl are aspreviously described. Preferred alkylaryls comprise a lower alkyl group.Non-limiting example of a suitable alkylaryl group is tolyl. The bond tothe parent moiety is through the aryl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring systemcomprising about 3 to about 10 carbon atoms, preferably about 5 to about10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7ring atoms. The cycloalkyl can be optionally substituted with one ormore “ring system substituents” which may be the same or different, andare as defined above. Non-limiting examples of suitable monocycliccycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyland the like. Non-limiting examples of suitable multicyclic cycloalkylsinclude 1-decalinyl, norbornyl, adamantyl and the like, as well aspartially saturated species such as, for example, indanyl,tetrahydronaphthyl and the like.

“Halogen” or “halo” means fluorine, chlorine, bromine, or iodine.Preferred are fluorine, chlorine and bromine.

“Ring system substituent” means a substituent attached to an aromatic ornon-aromatic ring system which, for example, replaces an availablehydrogen on the ring system. Ring system substituents may be the same ordifferent, each being independently selected from the group consistingof alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl,heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl,hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo,nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,cycloalkyl, heterocyclyl, —C(═N—CN)—NH₂, —C(═NH)—NH₂, —C(═NH)—NH(alkyl),Y₁Y₂N—, Y₁Y₂N-alkyl-, Y₁Y₂NC(O)—, Y₁Y₂NSO₂— and —SO₂NY₁Y₂, wherein Y₁and Y₂ can be the same or different and are independently selected fromthe group consisting of hydrogen, alkyl, aryl, cycloalkyl, and aralkyl.“Ring system substituent” may also mean a single moiety whichsimultaneously replaces two available hydrogens on two adjacent carbonatoms (one H on each carbon) on a ring system. Examples of such moietyare methylene dioxy, ethylenedioxy, —C(CH₃)₂— and the like which formmoieties such as, for example:

“Heterocyclyl” means a non-aromatic saturated monocyclic or multicyclicring system comprising about 3 to about 10 ring atoms, preferably about5 to about 10 ring atoms, in which one or more of the atoms in the ringsystem is an element other than carbon, for example nitrogen, oxygen orsulfur, alone or in combination. There are no adjacent oxygen and/orsulfur atoms present in the ring system. Preferred heterocyclyls containabout 5 to about 6 ring atoms. The prefix aza, oxa or thia before theheterocyclyl root name means that at least a nitrogen, oxygen or sulfuratom respectively is present as a ring atom. Any —NH in a heterocyclylring may exist protected such as, for example, as an —N(Boc), —N(CBz),—N(Tos) group and the like; such protections are also considered part ofthis invention. The heterocyclyl can be optionally substituted by one ormore “ring system substituents” which may be the same or different, andare as defined herein. The nitrogen or sulfur atom of the heterocyclylcan be optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclylrings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl,tetrahydrothiophenyl, lactam, lactone, and the like.

It should be noted that in hetero-atom containing ring systems of thisinvention, there are no hydroxyl groups on carbon atoms adjacent to a N,O or S, as well as there are no N or S groups on carbon adjacent toanother heteroatom. Thus, for example, in the ring:

there is no —OH attached directly to carbons marked 2 and 5.

It should also be noted that tautomeric forms such as, for example, themoieties:

are considered equivalent in certain embodiments of this invention.

“Alkynylalkyl” means an alkynyl-alkyl-group in which the alkynyl andalkyl are as previously described. Preferred alkynylalkyls contain alower alkynyl and a lower alkyl group. The bond to the parent moiety isthrough the alkyl. Non-limiting examples of suitable alkynylalkyl groupsinclude propargylmethyl.

“Heteroaralkyl” means a heteroaryl-alkyl-group in which the heteroaryland alkyl are as previously described. Preferred heteroaralkyls containa lower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude pyridylmethyl, and quinolin-3-ylmethyl. The bond to the parentmoiety is through the alkyl.

“Hydroxyalkyl” means a HO-alkyl-group in which alkyl is as previouslydefined. Preferred hydroxyalkyls contain lower alkyl. Non-limitingexamples of suitable hydroxyalkyl groups include hydroxymethyl and2-hydroxyethyl.

“Acyl” means an H—C(O)—, alkyl-C(O)— or cycloalkyl-C(O)—, group in whichthe various groups are as previously described. The bond to the parentmoiety is through the carbonyl. Preferred acyls contain a lower alkyl.Non-limiting examples of suitable acyl groups include formyl, acetyl andpropanoyl.

“Aroyl” means an aryl-C(O)— group in which the aryl group is aspreviously described. The bond to the parent moiety is through thecarbonyl. Non-limiting examples of suitable groups include benzoyl and1-naphthoyl.

“Alkoxy” means an alkyl-O— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkoxy groupsinclude methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond tothe parent moiety is through the ether oxygen.

“Aryloxy” means an aryl-O— group in which the aryl group is aspreviously described. Non-limiting examples of suitable aryloxy groupsinclude phenoxy and naphthoxy. The bond to the parent moiety is throughthe ether oxygen.

“Aralkyloxy” means an aralkyl-O— group in which the aralkyl group is aspreviously described. Non-limiting examples of suitable aralkyloxygroups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to theparent moiety is through the ether oxygen.

“Alkylthio” means an alkyl-S— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkylthio groupsinclude methylthio and ethylthio. The bond to the parent moiety isthrough the sulfur.

“Arylthio” means an aryl-S— group in which the aryl group is aspreviously described. Non-limiting examples of suitable arylthio groupsinclude phenylthio and naphthylthio. The bond to the parent moiety isthrough the sulfur.

“Aralkylthio” means an aralkyl-S— group in which the aralkyl group is aspreviously described. Non-limiting example of a suitable aralkylthiogroup is benzylthio. The bond to the parent moiety is through thesulfur.

“Alkoxycarbonyl” means an alkyl-O—CO— group. Non-limiting examples ofsuitable alkoxycarbonyl groups include methoxycarbonyl andethoxycarbonyl. The bond to the parent moiety is through the carbonyl.

“Aryloxycarbonyl” means an aryl-O—C(O)— group. Non-limiting examples ofsuitable aryloxycarbonyl groups include phenoxycarbonyl andnaphthoxycarbonyl. The bond to the parent moiety is through thecarbonyl.

“Aralkoxycarbonyl” means an aralkyl-O—C(O)— group. Non-limiting exampleof a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond tothe parent moiety is through the carbonyl.

“Alkylsulfonyl” means an alkyl-S(O₂)— group. Preferred groups are thosein which the alkyl group is lower alkyl. The bond to the parent moietyis through the sulfonyl.

“Arylsulfonyl” means an aryl-S(O₂)— group. The bond to the parent moietyis through the sulfonyl.

The term “substituted” means that one or more hydrogens on thedesignated atom is replaced with a selection from the indicated group,provided that the designated atom's normal valency under the existingcircumstances is not exceeded, and that the substitution results in astable compound. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds. By“stable compound” or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “one or more” or “at least one”, when indicating the number ofsubstituents, compounds, combination agents and the like, refers to atleast one, and up to the maximum number of chemically and physicallypermissible, substituents, compounds, combination agents and the like,that are present or added, depending on the context. Such techniques andknowledge are well known within the skills of the concerned artisan.

The term “optionally substituted” means optional substitution with thespecified groups, radicals or moieties.

The term “isolated” or “in isolated form” for a compound refers to thephysical state of said compound after being isolated from a syntheticprocess or natural source or combination thereof. The term “purified” or“in purified form” for a compound refers to the physical state of saidcompound after being obtained from a purification process or processesdescribed herein or well known to the skilled artisan, in sufficientpurity to be characterizable by standard analytical techniques describedherein or well known to the skilled artisan.

It should also be noted that any heteroatom with unsatisfied valences inthe text, schemes, examples and Tables herein is assumed to have thehydrogen atom(s) to satisfy the valences.

When a functional group in a compound is termed “protected”, this meansthat the group is in modified form to preclude undesired side reactionsat the protected site when the compound is subjected to a reaction.Suitable protecting groups will be recognized by those with ordinaryskill in the art as well as by reference to standard textbooks such as,for example, T. W. Greene et al, Protective Groups in organic Synthesis(1991), Wiley, New York.

When any variable (e.g., aryl, heterocycle, R², etc.) occurs more thanone time in any constituent or in Formula 1, its definition on eachoccurrence is independent of its definition at every other occurrence.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are alsocontemplated herein. The term “prodrug”, as employed herein, denotes acompound that is a drug precursor which, upon administration to asubject, undergoes chemical conversion by metabolic or chemicalprocesses to yield a compound of Formula 1 or a salt and/or solvatethereof. A discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, (1987)Edward B. Roche, ed., American Pharmaceutical Association and PergamonPress, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this inventionwith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the present inventioneffective in inhibiting the CDK(s) and thus producing the desiredtherapeutic, ameliorative, inhibitory or preventative effect.

The compounds of Formula 1 can form salts which are also within thescope of this invention. Reference to a compound of Formula 1 herein isunderstood to include reference to salts thereof, unless otherwiseindicated. The term “salt(s)”, as employed herein, denotes acidic saltsformed with inorganic and/or organic acids, as well as basic saltsformed with inorganic and/or organic bases. In addition, when a compoundof Formula 1 contains both a basic moiety, such as, but not limited to apyridine or imidazole, and an acidic moiety, such as, but not limited toa carboxylic acid, zwitterions (“inner salts”) may be formed and areincluded within the term “salt(s)” as used herein. Pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts arepreferred, although other salts are also useful. Salts of the compoundsof the Formula 1 may be formed, for example, by reacting a compound ofFormula 1 with an amount of acid or base, such as an equivalent amount,in a medium such as one in which the salt precipitates or in an aqueousmedium followed by lyophilization.

Exemplary acid addition salts include acetates, ascorbates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, fumarates, hydrochlorides,hydrobromides, hydroiodides, lactates, maleates, methanesulfonates,naphthalenesulfonates, nitrates, oxalates, phosphates, propionates,salicylates, succinates, sulfates, tartarates, thiocyanates,toluenesulfonates (also known as tosylates) and the like. Additionally,acids which are generally considered suitable for the formation ofpharmaceutically useful salts from basic pharmaceutical compounds arediscussed, for example, by P. Stahl et al, Camille G. (eds.) Handbook ofPharmaceutical Salts. Properties, Selection and Use. (2002) Zurich:Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977)66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33201-217; Anderson et al, The Practice of Medicinal Chemistry (1996),Academic Press, New York; and in The Orange Book (Food & DrugAdministration, Washington, D.C. on their website). These disclosuresare incorporated herein by reference thereto.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salts with organic bases (for example,organic amines) such as dicyclohexylamines, t-butyl amines, and saltswith amino acids such as arginine, lysine and the like. Basicnitrogen-containing groups may be quarternized with agents such as loweralkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides andiodides), dialkyl sulfates (e.g. dimethyl, diethyl, and dibutylsulfates), long chain halides (e.g. decyl, lauryl, and stearylchlorides, bromides and iodides), aralkyl halides (e.g. benzyl andphenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Pharmaceutically acceptable esters of the present compounds include thefollowing groups: (1) carboxylic acid esters obtained by esterificationof the hydroxy groups, in which the non-carbonyl moiety of thecarboxylic acid portion of the ester grouping is selected from straightor branched chain alkyl (for example, acetyl, n-propyl, t-butyl, orn-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (forexample, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (forexample, phenyl optionally substituted with, for example, halogen,C₁₋₄alkyl, or C₁₋₄alkoxy or amino); (2) sulfonate esters, such as alkyl-or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters(for example, L-valyl or L-isoleucyl); (4) phosphonate esters and (5)mono-, di- or triphosphate esters. The phosphate esters may be furtheresterified by, for example, a C₁₋₂₀ alcohol or reactive derivativethereof, or by a 2,3-di(C₆₋₂₄)acyl glycerol.

Compounds of Formula 1, and salts, solvates and prodrugs thereof, mayexist in their tautomeric form (for example, as an amide or iminoether). All such tautomeric forms are contemplated herein as part of thepresent invention.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the present compounds (including those of the salts,solvates and prodrugs of the compounds as well as the salts and solvatesof the prodrugs), such as those which may exist due to asymmetriccarbons on various substituents, including enantiomeric forms (which mayexist even in the absence of asymmetric carbons), rotameric forms,atropisomers, and diastereomeric forms, are contemplated within thescope of this invention, as are positional isomers (such as, forexample, 4-pyridyl and 3-pyridyl). Individual stereoisomers of thecompounds of the invention may, for example, be substantially free ofother isomers, or may be admixed, for example, as racemates or with allother, or other selected, stereoisomers. The chiral centers of thepresent invention can have the S or R configuration as defined by theIUPAC 1974 Recommendations. The use of the terms “salt”, “solvate”“prodrug” and the like, is intended to equally apply to the salt,solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers,positional isomers, racemates or prodrugs of the inventive compounds.

Polymorphic forms of the compounds of Formula I, and of the salts,solvates and prodrugs of the compounds of Formula I, are intended to beincluded in the present invention.

It is to be understood that the utility of the compounds of Formula 1for the therapeutic applications discussed herein is applicable to eachcompound by itself or to the combination or combinations of one or morecompounds of Formula 1 as illustrated, for example, in the nextimmediate paragraph. The same understanding also applies topharmaceutical composition(s) comprising such compound or compounds andmethod(s) of treatment involving such compound or compounds.

The compounds according to the invention can have pharmacologicalproperties; in particular, the compounds of Formula 1 can be inhibitorsof HCV protease, each compound by itself or one or more compounds ofFormula 1 can be combined with one or more compounds selected fromwithin Formula 1. The compound(s) can be useful for treating diseasessuch as, for example, HCV, HIV, (AIDS, Acquired Immune DeficiencySyndrome), and related disorders, as well as for modulating the activityof hepatitis C virus (HCV) protease, preventing HCV, or ameliorating oneor more symptoms of hepatitis C.

The compounds of Formula 1 may be used for the manufacture of amedicament to treat disorders associated with the HCV protease, forexample, the method comprising bringing into intimate contact a compoundof Formula 1 and a pharmaceutically acceptable carrier.

In another embodiment, this invention provides pharmaceuticalcompositions comprising the inventive compound or compounds as an activeingredient. The pharmaceutical compositions generally additionallycomprise at least one pharmaceutically acceptable carrier diluent,excipient or carrier (collectively referred to herein as carriermaterials). Because of their HCV inhibitory activity, suchpharmaceutical compositions possess utility in treating hepatitis C andrelated disorders.

In yet another embodiment, the present invention discloses methods forpreparing pharmaceutical compositions comprising the inventive compoundsas an active ingredient. In the pharmaceutical compositions and methodsof the present invention, the active ingredients will typically beadministered in admixture with suitable carrier materials suitablyselected with respect to the intended form of administration, i.e. oraltablets, capsules (either solid-filled, semi-solid filled or liquidfilled), powders for constitution, oral gels, elixirs, dispersiblegranules, syrups, suspensions, and the like, and consistent withconventional pharmaceutical practices. For example, for oraladministration in the form of tablets or capsules, the active drugcomponent may be combined with any oral non-toxic pharmaceuticallyacceptable inert carrier, such as lactose, starch, sucrose, cellulose,magnesium stearate, dicalcium phosphate, calcium sulfate, talc,mannitol, ethyl alcohol (liquid forms) and the like. Moreover, whendesired or needed, suitable binders, lubricants, disintegrating agentsand coloring agents may also be incorporated in the mixture. Powders andtablets may be comprised of from about 5 to about 95 percent inventivecomposition.

Suitable binders include starch, gelatin, natural sugars, cornsweeteners, natural and synthetic gums such as acacia, sodium alginate,carboxymethylcellulose, polyethylene glycol and waxes. Among thelubricants there may be mentioned for use in these dosage forms, boricacid, sodium benzoate, sodium acetate, sodium chloride, and the like.Disintegrants include starch, methylcellulose, guar gum and the like.

Sweetening and flavoring agents and preservatives may also be includedwhere appropriate. Some of the terms noted above, namely disintegrants,diluents, lubricants, binders and the like, are discussed in more detailbelow.

Additionally, the compositions of the present invention may beformulated in sustained release form to provide the rate controlledrelease of any one or more of the components or active ingredients tooptimize the therapeutic effects, i.e. HCV inhibitory activity and thelike. Suitable dosage forms for sustained release include layeredtablets containing layers of varying disintegration rates or controlledrelease polymeric matrices impregnated with the active components andshaped in tablet form or capsules containing such impregnated orencapsulated porous polymeric matrices.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injections or addition of sweeteners and pacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier such as inert compressed gas, e.g.nitrogen.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides such as cocoa butter is first melted, and theactive ingredient is dispersed homogeneously therein by stirring orsimilar mixing. The molten homogeneous mixture is then poured intoconvenient sized molds, allowed to cool and thereby solidify.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions may take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

The compounds of the invention may also be administered orally,intravenously, intranasally or subcutaneously.

The compounds of the invention may also comprise preparations which arein a unit dosage form. In such form, the preparation is subdivided intosuitably sized unit doses containing appropriate quantities of theactive components, e.g., an effective amount to achieve the desiredpurpose.

The quantity of the inventive active composition in a unit dose ofpreparation may be generally varied or adjusted from about 1.0 milligramto about 1,000 milligrams, preferably from about 1.0 to about 950milligrams, more preferably from about 1.0 to about 500 milligrams, andtypically from about 1 to about 250 milligrams, according to theparticular application. The actual dosage employed may be varieddepending upon the patient's age, sex, weight and severity of thecondition being treated. Such techniques are well known to those skilledin the art.

Generally, the human oral dosage form containing the active ingredientscan be administered 1 or 2 times per day. The amount and frequency ofthe administration will be regulated according to the judgment of theattending clinician. A generally recommended daily dosage regimen fororal administration may range from about 1.0 milligram to about 1,000milligrams per day, in single or divided doses.

Some useful terms are described below:

Capsule—refers to a special container or enclosure made of methylcellulose, polyvinyl alcohols, or denatured gelatins or starch forholding or containing compositions comprising the active ingredients.Hard shell capsules are typically made of blends of relatively high gelstrength bone and pork skin gelatins. The capsule itself may containsmall amounts of dyes, opaquing agents, plasticizers and preservatives.

Tablet—refers to a compressed or molded solid dosage form containing theactive ingredients with suitable diluents. The tablet can be prepared bycompression of mixtures or granulations obtained by wet granulation, drygranulation or by compaction.

Oral gel—refers to the active ingredients dispersed or solubilized in ahydrophillic semi-solid matrix.

Powder for constitution refers to powder blends containing the activeingredients and suitable diluents which can be suspended in water orjuices.

Diluent—refers to substances that usually make up the major portion ofthe composition or dosage form. Suitable diluents include sugars such aslactose, sucrose, mannitol and sorbitol; starches derived from wheat,corn, rice and potato; and celluloses such as microcrystallinecellulose. The amount of diluent in the composition can range from about10 to about 90% by weight of the total composition, preferably fromabout 25 to about 75%, more preferably from about 30 to about 60% byweight, even more preferably from about 12 to about 60%.

Disintegrant—refers to materials added to the composition to help itbreak apart (disintegrate) and release the medicaments. Suitabledisintegrants include starches; “cold water soluble” modified starchessuch as sodium carboxymethyl starch; natural and synthetic gums such aslocust bean, karaya, guar, tragacanth and agar; cellulose derivativessuch as methylcellulose and sodium carboxymethylcellulose;microcrystalline celluloses and cross-linked microcrystalline cellulosessuch as sodium croscarmellose; alginates such as alginic acid and sodiumalginate; clays such as bentonites; and effervescent mixtures. Theamount of disintegrant in the composition can range from about 2 toabout 15% by weight of the composition, more preferably from about 4 toabout 10% by weight.

Binder—refers to substances that bind or “glue” powders together andmake them cohesive by forming granules, thus serving as the “adhesive”in the formulation. Binders add cohesive strength already available inthe diluent or bulking agent. Suitable binders include sugars such assucrose; starches derived from wheat, corn rice and potato; natural gumssuch as acacia, gelatin and tragacanth; derivatives of seaweed such asalginic acid, sodium alginate and ammonium calcium alginate; cellulosicmaterials such as methylcellulose and sodium carboxymethylcellulose andhydroxypropylmethylcellulose; polyvinylpyrrolidone; and inorganics suchas magnesium aluminum silicate. The amount of binder in the compositioncan range from about 2 to about 20% by weight of the composition, morepreferably from about 3 to about 10% by weight, even more preferablyfrom about 3 to about 6% by weight.

Lubricant—refers to a substance added to the dosage form to enable thetablet, granules, etc. after it has been compressed, to release from themold or die by reducing friction or wear. Suitable lubricants includemetallic stearates such as magnesium stearate, calcium stearate orpotassium stearate; stearic acid; high melting point waxes; and watersoluble lubricants such as sodium chloride, sodium benzoate, sodiumacetate, sodium oleate, polyethylene glycols and d′l-leucine. Lubricantsare usually added at the very last step before compression, since theymust be present on the surfaces of the granules and in between them andthe parts of the tablet press. The amount of lubricant in thecomposition can range from about 0.2 to about 5% by weight of thecomposition, preferably from about 0.5 to about 2%, more preferably fromabout 0.3 to about 1.5% by weight.

Glident—material that prevents caking and improve the flowcharacteristics of granulations, so that flow is smooth and uniform.Suitable glidents include silicon dioxide and talc. The amount ofglident in the composition can range from about 0.1% to about 5% byweight of the total composition, preferably from about 0.5 to about 2%by weight.

Coloring agents—excipients that provide coloration to the composition orthe dosage form. Such excipients can include food grade dyes and foodgrade dyes adsorbed onto a suitable adsorbent such as clay or aluminumoxide. The amount of the coloring agent can vary from about 0.1 to about5% by weight of the composition, preferably from about 0.1 to about 1%.

Bioavailability—refers to the rate and extent to which the active drugingredient or therapeutic moiety is absorbed into the systemiccirculation from an administered dosage form as compared to a standardor control.

Conventional methods for preparing tablets are known. Such methodsinclude dry methods such as direct compression and compression ofgranulation produced by compaction, or wet methods or other specialprocedures. Conventional methods for making other forms foradministration such as, for example, capsules, suppositories and thelike are also well known.

Another embodiment of the invention discloses the use of the inventivecompounds or pharmaceutical compositions disclosed above for treatmentof diseases such as, for example, hepatitis C and the like. The methodcomprises administering a therapeutically effective amount of theinventive compound or pharmaceutical composition to a patient havingsuch a disease or diseases and in need of such a treatment.

In yet another embodiment, the compounds of the invention may be usedfor the treatment of HCV in humans in monotherapy mode or in acombination therapy (e.g., dual combination, triple combination etc.)mode such as, for example, in combination with antiviral and/orimmunomodulatory agents. Examples of such antiviral and/orimmunomodulatory agents include Ribavirin (from Schering-PloughCorporation, Madison, N.J.) and Levovirin™ (from ICN Pharmaceuticals,Costa Mesa, Calif.), VP 50406™ (from Viropharma, Incorporated, Exton,Pa.), ISIS 14803™ (from ISIS Pharmaceuticals, Carlsbad, Calif.),Heptazyme™ (from Ribozyme Pharmaceuticals, Boulder, Colo.), VX 497™(from Vertex Pharmaceuticals, Cambridge, Mass.), Thymosin™ (fromSciClone Pharmaceuticals, San Mateo, Calif.), Maxamine™ (MaximPharmaceuticals, San Diego, Calif.), mycophenolate mofetil (fromHoffman-LaRoche, Nutley, N.J.), interferon (such as, for example,interferon-alpha, PEG-interferon alpha conjugates) and the like.“PEG-interferon alpha conjugates” are interferon alpha moleculescovalently attached to a PEG molecule. Illustrative PEG-interferon alphaconjugates include interferon alpha-2a (Roferon™, from Hoffman La-Roche,Nutley, N.J.) in the form of pegylated interferon alpha-2a (e.g., assold under the trade name Pegasys™), interferon alpha-2b (Intron™, fromSchering-Plough Corporation) in the form of pegylated interferonalpha-2b (e.g., as sold under the trade name PEG-Intron™), interferonalpha-2c (Berofor Alpha™, from Boehringer Ingelheim, Ingelheim, Germany)or consensus interferon as defined by determination of a consensussequence of naturally occurring interferon alphas (Infergen™, fromAmgen, Thousand Oaks, Calif.).

As stated earlier, the invention includes tautomers, rotamers,enantiomers and other stereoisomers of the inventive compounds also.Thus, as one skilled in the art appreciates, some of the inventivecompounds may exist in suitable isomeric forms. Such variations arecontemplated to be within the scope of the invention.

Another embodiment of the invention discloses a method of making thecompounds disclosed herein. The compounds may be prepared by severaltechniques known in the art. Illustrative procedures are outlined in thefollowing reaction schemes. The illustrations should not be construed tolimit the scope of the invention which is defined in the appendedclaims. Alternative mechanistic pathways and analogous structures willbe apparent to those skilled in the art.

It is to be understood that while the following illustrative schemesdescribe the preparation of a few representative inventive compounds,suitable substitution of any of both the natural and unnatural aminoacids will result in the formation of the desired compounds based onsuch substitution. Such variations are contemplated to be within thescope of the invention.

For the procedures described below, the following abbreviations areused:

Abbreviations

Abbreviations which are used in the descriptions of the schemes,preparations and the examples that follow are:

THF: Tetrahydrofuran

DMF: N,N-Dimethylformamide

EtOAc: Ethyl acetate

AcOH: Acetic acid

HOOBt: 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one

EDCl: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

NMM: N-Methylmorpholine

ADDP: 1,1′-(Azodicarbobyl)dipiperidine

DEAD: Diethylazodicarboxylate

MeOH: Methanol

EtOH: Ethanol

Et₂O: Diethyl ether

DMSO: Dimethylsulfoxide

HOBt: N-Hydroxybenzotriazole

PyBrOP: Bromo-tris-pyrrolidinophosphonium hexafluorophosphate

DCM: Dichloromethane

DCC: 1,3-Dicyclohexylcarbodiimide

TEMPO: 2,2,6,6-Tetramethyl-1-piperidinyloxy

Phg: Phenylglycine

Chg: Cyclohexylglycine

Bn: Benzyl

Bzl: Benzyl

Et: Ethyl

Ph: Phenyl

iBoc: isobutoxycarbonyl

iPr: isopropyl

^(t)Bu or Bu^(t): tert-Butyl

Boc: tert-Butyloxycarbonyl

Cbz: Benzyloxycarbonyl

Cp: Cylcopentyldienyl

Ts: p-toluenesulfonyl

MCPBA: 3-chloroperbenzoic acid.

Me: Methyl

HATU: O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate

DMAP: 4-N,N-Dimethylaminopyridine

Bop: Benzotriazol-1-yl-oxy-tris(dimethylamino)hexafluorophosphate

PCC: Pyridiniumchlorochromate

Other abbreviations are commonly used abbreviations Such as according tothe guidelines published by Journal of Organic Chemistry.

General Schemes for Preparation of Target Compounds

Compounds of the present invention were synthesized using the generalschemes (Methods A-E) described below.

Method A

Deprotection of the N-Boc functionality of 1.01 under acidic conditionsprovided the hydrochloride salt 1.02 which was subsequently coupled withN-Boc-tert-leucine under peptide coupling methodology (Louis A Carpinoet al. “Preparation of uronium and immonium salts for peptide coupling”,WO 2002094822, pp. 76) to afford 1.03. N-Boc deprotection followed bytreatment with appropriate isocyanate gave the urea 1.05. Hydrolysis ofthe methyl ester provided the acid 1.06. Peptide coupling of the acid1.06 with the appropriate P₁—P′ primary amide moiety afforded thehydroxyl amide 1.07. Oxidation (Moffatt, or Dess-Martin's) resulted inthe target compound 1.08.

Method BPeptide coupling of the acid 1.06 with the appropriate P₁—P′ secondaryamide moiety afforded the hydroxyl amide 1.09. Oxidation (Moffatt orDess-Martin's) resulted in the target compound 1.10.Method CIn another variation, peptide coupling of the N-Boc-P2-P₃-acid 1.03 withthe appropriate P₁—P′ amide moiety afforded the hydroxyl amide 1.11.Oxidation (Moffatt or Dess-Martin's) resulted in the keto-amide 1.12.Deprotection of the N-Boc using either formic acid or 4 M HCl in dioxanegave the formate or hydrochloride salt 1.13. Treatment with a suitableisocyanate (or isocyanate equivalent) resulted in the target compound1.14.

Method DIn yet another variation, the hydrochloride salt 1.13 was converted tothe 4-nitrophenyl carbamate 1.15 by reaction with 4-nitrophenylchloroformate. Subsequent treatment with an amine (or aminehydrochloride salt) of choice provided the target compound 1.14.

Method EIn yet another variation, the dipeptide hydrochloride salt 1.04 wasconverted to the 4-nitrophenyl carbamate as described above. Treatmentwith an amine (or amine hydrochloride salt) of choice provided the ureaderivative 1.05. Hydrolysis and further elaboration as described inMethods A/B provided the target compounds 1.14.

Preparation of IntermediatesPreparation of P1-P′ moietiesPreparation of Intermediates 10.11 and 10.12Step 1

A stirred solution of ketimine 10.01 (50 g, 187.1 mmol) under N₂ in dryTHF (400 mL) was cooled to −78° C. and treated with 1 M solution ofK-^(t)BuO (220 mL, 1.15 equiv.) in THF. The reaction mixture was warmedto 0° C. and stirred for 1 h and treated with bromomethyl cyclobutane(28 mL, 249 mmol). The reaction mixture was stirred at room temperaturefor 48 h and concentrated in vacuo. The residue was dissolved in Et₂O(300 mL) and treated with aq. HCl (2 M, 300 mL) The resulting solutionwas stirred at room temperature for 5 h and extracted with Et₂O (1 L).The aqueous layer was made basic to pH ˜12-14 with NaOH (50% aq.) andextracted with CH₂Cl₂ (3×300 mL). The combined organic layers were dried(MgSO₄), filtered, and concentrated to give the pure amine (10.02, 18 g)as a colorless oil.

Step 2.

A solution of the amine 10.02 (18 g, 105.2 mmol) at 0° C. in CH₂Cl₂ (350mL) was treated with di-tert-butyldicarbonate (23 g, 105.4 mmol) andstirred at rt. for 12 h. After the completion of the reaction (TLC), thereaction mixture was concentrated in vacuo and the residue was dissolvedin THF/H₂O (200 ml, 1:1) and treated with LiOH.H₂O (6.5 g, 158.5 mmol)and stirred at room temperature for 3 h. The reaction mixture wasconcentrated and the basic aqueous layer was extracted with Et₂O. Theaqueous layer was acidified with conc. HCl to pH ˜1-2 and extracted withCH₂Cl₂. The combined organic layers were dried (MgSO₄), filtered, andconcentrated in vacuo to yield 10.03 as a colorless viscous oil whichwas used for the next step without any further purification.

Step 3.

A solution of the acid 10.03 (15.0 g, 62 mmol) in CH₂Cl₂ (250 mL) wastreated with BOP reagent (41.1 g, 93 mmol), N-methyl morpholine (27 mL),N,O-dimethyl hydroxylamine hydrochloride (9.07 g, 93 mmol) and stirredovernight at rt. The reaction mixture was diluted with 1 N aq. HCl (250mL), and the layers were separated and the aqueous layer was extractedwith CH₂Cl₂ (3×300 ml). The combined organic layers were dried (MgSO₄),filtered and concentrated in vacuo and purified by chromatography (SiO₂,EtOAc/Hex 2:3) to yield the amide 10.04 (15.0 g) as a colorless solid.

Step 4.

A solution of the amide 10.04 (15 g, 52.1 mmol) in dry THF (200 mL) wastreated dropwise with a solution of LiAlH₄ (1M, 93 mL, 93 mmol) at 0° C.The reaction mixture was stirred at room temperature for 1 h andcarefully quenched at 0° C. with a solution of KHSO₄ (10% aq.) andstirred for 0.5 h. The reaction mixture was diluted with aq. HCl (1 M,150 mL) and extracted with CH₂Cl₂ (3×200 mL), The combined organiclayers were washed with aq. HCl (1 M), saturated NaHCO₃, brine, anddried (MgSO₄). The mixture was filtered and concentrated in vacuo toyield 10.05 as a viscous colorless oil (14 g).

Step 5.

A solution of the aldehyde 10.05 (14 g, 61.6 mmol) in CH₂Cl₂ (50 mL),was treated with Et₃N (10.73 mL, 74.4 mmol), and acetone cyanohydrin(10.86 g, 127.57 mmol) and stirred at room temperature for 24 hrs. Thereaction mixture was concentrated in vacuo and diluted with aq. HCl (1M, 200 mL) and extracted into CH₂Cl₂ (3×200 mL). The combined organiclayer were washed with H₂O, brine, dried (MgSO₄), filtered, concentratedin vacuo and purified by chromatography (SiO₂, EtOAc/Hex 1:4) to yield10.06 (10.3 g) as a colorless liquid

Step 6.

Methanol saturated with HCl*, prepared by bubbling HCl gas through CH₃OH(700 ml) at 0° C., was treated with the cyanohydrin 10.06 and heated toreflux for 24 h. The reaction was concentrated in vacuo to yield 10.07,which was used in the next step without purification. *Alternatively 6MHCl prepared by addition of AcCl to dry methanol can also be used.

Step 7.

A solution of the amine hydrochloride 10.07 in CH₂Cl₂ (200 mL) wastreated with Et₃N (45.0 mL, 315 mmol) and Boc₂O (45.7 g, 209 mmol) at−78° C. The reaction mixture was then stirred at room temperatureovernight and diluted with HCl (2 M, 200 mL) and extracted into CH₂Cl₂.The combined organic layers were dried (MgSO₄) filtered, concentrated invacuo and purified by chromatography (EtOAc/Hex 1:4) to yield hydroxyester 10.08.

Step 8.

A solution of methyl ester 10.08 (3 g, 10.5 mmol) in THF/H₂O (1:1) wastreated with LiOH.H₂O (645 mg, 15.75 mmol) and stirred at rt. for 2 h.The reaction mixture was acidified with aq HCl (1 M, 15 mL) andconcentrated in vacuo. The residue was dried in vacuum to afford 10.09in quantitative yield.

Step 9

A solution of the acid 10.09 (from above) in CH₂Cl₂ (50 mL) and DMF (25mL) was treated with NH₄Cl (2.94 g, 55.5 mmol), EDCl (3.15 g, 16.5mmol), HOOBt (2.69 g, 16.5 mmol), and NMM (4.4 g, 44 mmol). The reactionmixture was stirred at room temperature for 3 d. The solvents wereremoved under vacuo and the residue was diluted with aq. HCl (250 mL)and extracted with CH₂Cl₂. The combined organic layers were washed withaq. Sat'd. NaHCO₃, dried (MgSO₄) filtered concentrated in vacuo toobtain 10.10, which was used as it was in the following steps.(Alternatively 10.10 can also be obtained directly by the reaction of10.06 (4.5 g, 17.7 mmol) with aq. H₂O₂ (10 mL), LiOH.H₂O (820 mg, 20.8mmol) at 0° C. in 50 mL of CH₃OH for 0.5 h.)

Step 10.

A solution of 10.10 obtained in the previous step was dissolved in 4 NHCl in dioxane and stirred at rt. for 2 h. The reaction mixture wasconcentrated in vacuo to give the intermediate 10.11 as a solid, whichwas used without further purification.

Step 11.

The required intermediate 10.12 was obtained from compound 10.09 usingessentially the procedures described above in Steps 9, 10 using 2.0equivalents of allylamine instead of ammonium chloride.

Preparation of Intermediate 11.01

Step 1

To a solution of 4-pentyn-1-ol, 11.02 (4.15 g; Aldrich) was addedDess-Martin Periodinane (30.25 g; Aldrich) and the resulting mixture wasstirred for 45 min. before the addition of(tert-Butoxycarbonylmethylene)triphenylphosphorane (26.75 g; Aldrich).The resulting dark reaction was stirred overnight, diluted with EtOAc),washed with aq. sodium sulfite. sat. aq. NaHCO3, water, brine and dried.The volatiles were removed under reduced pressure and the residue waspurified by silica gel column chromatography using 1% EtOAc in hexanesas eluent to give the desired compound, 11.03 (3.92 g). Some impurefractions were also obtained but set aside at this time.Step 2

Using the alkene 11.03 (1.9 g) in n-propanol (20 ml; Aldrich)), benzylcarbamate (4.95 g; Aldrich) in n-propanol (40 ml), NaOH (1.29 g) inwater (79 ml), tert-butyl hypochlorite (3.7 ml), (DHQ)2PHAL (0.423 g;Aldrich)) in n-propanol (37.5 ml), and potassium osmate:dehydrate(0.1544 g; Aldrich) and the procedure set forth in Angew. Chem. Int. Ed.Engl (1998), 35, (23/24), pp. 2813-7 gave a crude product which waspurified by silica gel column chromatography using EtOAc:Hexanes (1:5)to give the desired amino alcohol 11.04 (1.37 g, 37%) as a white solid.Step 3

To the ester 11.04 (0.700 g) was added 4M HCl in dioxane (20 ml;Aldrich) and the resulting mixture was allowed to stand at roomtemperature overnight. The volatiles were removed under reduced pressureto give the acid 11.05 (0.621 g) as a white solid.Step 4

BOP reagent (3.65 g; Sigma) followed by triethylamine (3.45 ml) wereadded to a dichloromethane (20 ml) solution of the carboxylic acid 11.05(2.00 g) and allyl amine (0.616 ml) at room temperature and theresulting mixture was stirred overnight. The reaction mixture waspartitioned between EtOAc and 10% aq. HCl. The organic phase wasseparated, washed with sat. aq. sodium bicarbonate, water, dried(magnesium sulfate). The crude reaction product was purified by silicagel column chromatography using (EtOAc:Hexanes; 70:30) as eluent toprovide the desired amide 11.01 (1.73 g) as a viscous yellow oil.Preparation of Intermediates 12.03 and 12.04Step 1

Compound 12.01 (Compound 12.01 was obtained commercially or can besynthesized using similar chemistry as outlined for the synthesis of10.11 using bromomethyl cyclopropane instead of bromomethylcyclobutane)was converted to the required material 12.02 using essentially theprocedures described for Intermediate 10.11, Steps 3-8.Step 2

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.11, Steps 9,10.Step 3

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.12, Step 11.Preparation of Intermediate 13.01Step 1

To a stirred solution of 1-nitrobutane, 13.02 (16.5 g, 0.16 mol) andglyoxylic acid in H₂O (28.1 g, 0.305 mol) and MeOH (122 mL) at 0° C.-5°C., was added dropwise triethylamine (93 mL, 0.667 mol) over 2 hrs. Thesolution was warmed to room temperature, stirred overnight andconcentrated to dryness to give an oil. The oil was then dissolved inH₂O and acidified to pH=1 with 10% HCl, followed by extraction withEtOAc. The combined organic solution was washed with brine, dried overNa₂SO₄, filtered and concentrated to dryness to give the product 13.03(28.1 g, 99% yield).

Step 2

To a stirred solution of compound 13.03 (240 g, 1.35 mol) in acetic acid(1.25 L) was added 10% Pd/C (37 g). The resulting solution washydrogenated at 59 psi for 3 hrs and then at 60 psi overnight. Theacetic acid was then evaporated and azeotroped 3 times with toluene,then triturated with MeOH and ether. The solution was then filtered andazeotroped twice with toluene to afford 13.04 as an off white solid (131g, 0.891 mol, 66%).

Step 3

To a stirred solution of the amino acid 13.04 (2.0 g, 13.6 mmol) indioxane (10 mL) and H₂O (5 mL) at 0° C., was added 1N NaOH solution (4.3mL, 14.0 mmol). The resulting solution was stirred for 10 minutes,followed by addition of di-t-butyldicarbonate (0.110 g, 14.0 mmol) andstirred at 0° C. for 15 minutes. The solution was then warmed to roomtemperature, stirred for 45 minutes and kept at refrigerator overnightand concentrated to dryness to give a crude material. To the solution ofthis crude material in EtOAc (100 mL) and ice, was added KHSO₄ (3.36 g)and H₂O (32 mL) and stirred for 4-6 minutes. The organic layer was thenseparated and the aqueous layer was extracted twice with EtOAc and thecombined organic layer was washed with water, brine, dried over Na₂SO₄,filtered and concentrated to dryness to give the product 13.05 as aclear gum (3.0 g, 89% yield).

Step 4

Compound 13.05 was converted to the required intermediate 13.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 14.01

Step 1

Compound 14.02 was converted to the required material 14.03 usingessentially the procedures described for Intermediate 13.01, Steps 1-3.

Step 2

Compound 14.03 was converted to the required intermediate 14.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 15.01

Step 1

To a suspension of silver nitrite (9 g, 58.5 mmol) in diethyl ether (25mL) at 0° C. was added a solution of 4-iodo-1,1,1-trifluorobutane, 15.02(10 g, 42.0 mmol) in diethyl ether (25 mL) slowly through an additionfunnel (approx. 15 min). The resulting mixture was vigorously stirred at0° C. and warmed to rt. After 50 h, the solid material was filtered offthrough a celite pad. The resulting diethyl ether solution wasconcentrated in vacuo to give 15.03 as colorless oil, which was usedwithout further purification.

Step 2

Compound 15.03 was converted to the required material 15.04 usingessentially the procedures described for Intermediate 13.01, Steps 1-3.Step 3

Compound 15.04 was converted to the required intermediate 15.01 usingessentially the procedures described for Intermediate 10.12, Step 11.Preparation of Intermediate 16.01

The acid 16.02 (Winkler, D.; Burger, K., Synthesis, 1996, 1419) isprocessed as described above (preparation of Intermediate 10.12) to givethe expected intermediate 16.01.Preparation of P2/P3-P2 MoietiesPreparation of Intermediate 20.01

The amino ester 20.01 was prepared following the method of R. Zhang andJ. S. Madalengoitia (J. Org. Chem. 1999, 64, 330), with the exceptionthat the Boc group was cleaved by the reaction of the Boc-protectedamino acid with methanolic HCl. (Note: In a variation of the reportedsynthesis, the sulfonium ylide used for the construction of3,4-dimethylcyclopropyl ring was replaced with the correspondingphosphonium ylide)Preparation of Intermediate 21.01Step 1:

To a stirred solution of N-Boc-3,4-dehydroproline 21.02 (5.0 g, 23.5mmol), di-tert-butyl dicarbonate (7.5 g, 34.4 mmol), and4-N,N-dimethylaminopyridine (0.40 g, 3.33 mmol) in acetonitrile (100 mL)at room temperature was added triethylamine (5.0 mL, 35.6 mmol). Theresulting solution was stirred at this temperature for 18 h before itwas concentrated in vacuo. The dark brown residue was purified by flashcolumn chromatography eluting with 10-25% EtOAc/hexane to give theproduct 21.03 as a pale yellow oil (5.29 g, 84%).

Step 2:

To a stirred solution of the dehydroproline derivative 21.03 (10.1 g,37.4 mmol), benzyltriethylammonium chloride (1.60 g, 7.02 mmol) inchloroform (120 mL) at room temperature was added 50% aqueous sodiumhydroxide (120 g). After vigorously stirred at this temperature for 24h, the dark mixture was diluted with CH₂Cl₂ (200 mL) and diethyl ether(600 mL). After the layers were separated, the aqueous solution wasextracted with CH₂Cl₂/Et₂O (1:2, 3×600 mL). The organic solution wasdried (MgSO₄) and concentrated. The residue was purified by flash columnchromatography using 5-20% EtOAc/hexane to afford 9.34 g (71%) of 21.04as an off-white solid.

Step 3:

The solution of 21.04 (9.34 g, 26.5 mmol) in CH₂Cl₂ (25 mL) and CF₃CO₂H(50 mL) was stirred at room temperature for 4.5 h before it wasconcentrated in vacuo to give a brown residue, 21.05 which was used inStep 4 without further purification.

Step 4

Concentrated hydrochloric acid (4.5 mL) was added to a solution of theresidue 21.05 from Step 3 in methanol (70 mL) and the resulting mixturewas warmed to 65° C. in an oil bath. After 18 h, the mixture wasconcentrated in vacuo to give a brown oil 21.01, which was used furtherwithout purification.

Preparation of Intermediate 22.01

Step 1

Potassium bis(trimethylsilyl)amide (158 ml of a 0.5M solution intoluene; 79 mmol) was added to a stirred suspension ofcyclopropyltriphenylphosphonium bromide (33.12 g; 86.4 mmol) inanhydrous tetrahydrofuran (130 ml) and the resulting orange mixture wasstirred under an atmosphere of nitrogen at room temperature for a periodof 1 h., before the addition of the aldehyde 22.02 (9.68 g; 42.2 mmol)in THF (8 ml). The reaction was then refluxed under an atmosphere ofnitrogen for a period of 2 h. After cooling, methanol, diethyl ether andRochelles salt were added. The organic phase was separated, washed withbrine, dried and concentrated under reduced pressure. The crude reactionproduct was purified by silica gel column chromatography usingEtOAc-hexane (1:99) to EtOAc-hexane (5:95) to provide the alkene 22.03(8.47 g) as a yellow oil.

Step 2

A solution of 1M HCl in MeOH/MeOAc was prepared by adding 14.2 ml ofacetyl chloride dropwise into cold methanol and diluting the resultingsolution to 200 ml at room temperature. The carbamate 22.03 (9.49 g;37.5 mmol) was dissolved in methanol (12 ml) and added to 1M HCl inMeOH/MeOAc (150 ml) while cooled in an ice bath. The resulting mixturewas maintained at this temperature for 1 h., then the ice bath wasremoved and stirring continued overnight at room temperature. Thevolatiles were removed under reduced pressure to yield a yellow oilwhich was used in the next step without purification. The yellow oil wasdissolved in a mixture of THF (30 ml) and MeOH (20 ml) and treated withtriethylamine (15 ml; 108 mmol) until the solution was pH=9-10. Afterplacing in an ice bath, the mixture was treated with N-Boc-Gly-OSu(11.22 g; 41 mmol). The ice bath was withdrawn and the reaction stirredat room temp. for 1 h. The volatiles were removed under reduced pressureand the residue was purified by silica gel column chromatography usingmethanol (1-3%) in dichloromethane providing the desired amide 22.04(9.09 g).

Step 3

The alcohol 22.04 (9.09 g; 33.6 mmol) was dissolved in acetone (118.5ml) and treated with 2,2-dimethoxypropane (37.4 ml; 304 mmol) andBF₃:Et₂O (0.32 ml; 2.6 mmol) and the resulting mixture was stirred atroom temperature for a period of 5.5 h The reaction solution was treatedwith a few drops of triethylamine and the volatiles were removed underreduced pressure. The residue was purified by silica gel columnchromatography using 5-25% EtOAc in hexanes to provide the N,O-acetal22.05 (8.85 g).

Step 4

The carbamate 22.05 (8.81 g; 28.4 mmol) was dissolved in acetonitrile(45 ml) and the solution was cooled to −40° C. under an atmosphere ofnitrogen. Pyridine (6.9 ml; 85.3 mmol) followed by nitrosiumtetrafluoroborate (6.63 g; 56.8 mmol) were added and the resultingreaction mixture maintained below 0° C. until TLC indicated that nostarting material remained (approx. 2.25 h.). Pyrrolidine (20 ml; 240mmol) was added and the cooling bath was withdrawn and stirring wascontinued at room temperature for 1 h. and then the volatiles wereremoved under reduced pressure. The residue was quickly passed through apad of silica gel to provide a yellow oil. The yellow oil was dissolvedin anhydrous benzene (220 ml) and palladium acetate (0.317 g; 1.41 mmol)was added before heating the resulting mixture to reflux, under anatmosphere of nitrogen for a period of 1.5 h. After cooling, thevolatiles were removed under reduced pressure and the dark residue waspurified by silica gel column chromatography using EtOAc-hexane (1:4) toprovide the 1) the trans-pyrrolidinone 22.06 (1.94 g) followed by ii)the cis-pyrrolidinone 22.07 (1.97 g).

Step 5

Freshly prepared 1M HCl in MeOAc/MeOH (10 ml; as described above) wasadded to the N,O-acetal 22.06 and stirred at room temperature for 1 h.The solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography using 0-4% MeOH indichloromethane as eluent to provide the desired alcohol 22.08 (1.42 g),a yellow oil.

Step 6

To a solution of the lactam 22.08 (1.29 g; 8.44 mmol) in anhydroustetrahydrofuran (55 ml) was added lithium aluminum hydride (2.40 g; 63.2mmol) and the resulting mixture was refluxed for 8 h. After cooling,water, followed by 15% aq. NaOH were added and the resulting mixture wasfiltered through celite and the solid was washed thoroughly with THF andMeOH. The solvent was removed under reduced pressure and the residueredissolved in dichloromethane, dried and concentrated under reducedpressure to provide the pyrrolidine, used without purification. Hunigsbase (4.5 ml; 25.8 mmol) was added to a mixture of N-Boc-L-tert-Leu-OH(1.76 g; 7.6 mmol), The crude pyrrolidine and HATU (2.89 g; 7.6 mmol) inanhydrous dichloromethane (50 ml) at −60° C., under an atmosphere ofnitrogen. The resulting reaction was allowed to come to room temperatureslowly, overnight. EtOAc was added and the yellow solution was washedwith dil. aq. HCl, sat. aq. sodium bicarbonate, water, brine. Theorganic layer was dried and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography usingEtOAc:hexanes (1:3) to give the desired amide 22.09 (2.00 g).

Step 7

The alcohol 22.09 (2.00 g; 5.67 mmol) was dissolved in acetone (116 ml)and cooled in an ice bath for 10 min. This solution was then added to acooled Jones reagent (14.2 ml; approx 2 mmol/ml) and the resultingmixture was stirred at 5 C for 0.5 h and the cooling bath was removed.The reaction was stirred for a further 2 h. at room temp., before addingto sodium sulfate (28.54 g), celite (15 g) in EtOAc (100 ml).Isopropanol (15 ml) was added after 1 min and then stirred for a further10 min. and filtered. The filtrate was concentrated under reducedpressure, providing a brown oil which was dissolved in EtOAc. Thissolution was washed with water, 3% aq. citric acid, brine, dried andconcentrated to provide the desired carboxylic acid 22.01 (1.64 g) as awhite solid.

Preparation of Intermediate 23.01

Step 1:

To the mixture of ester 23.02 (6.0 g) and molecular sieve (5.2 g) inanhydrous methylene chloride (35 mL) was added pyrrolidine (5.7 mL,66.36 mmoL). The resulting brown slurry was stirred at room temperatureunder N₂ for 24 h, filtered and washed with anhydrous CH₃CN. Thecombined filtrate was concentrated to yield the desired product, 23.03.

Step 2:

To a solution of the product 23.03 from proceeding step in CH₃CN (35 mL)was added anhydrous K₂CO₃, methallyl chloride (2.77 g, 30.5 mmoL), NaI(1.07 g, 6.7 mmoL). The resulting slurry was stirred at ambienttemperature under N₂ for 24 h. 50 mL of ice-cold water was addedfollowed by 2N KHSO₄ solution until pH was 1. EtOAc (100 mL) was addedand the mixture was stirred for 0.75 h. Combined organic layer wascollected and washed with brine, dried over MgSO₄, and evaporated toyield the desired product, 23.04.

Step 3:

The product 23.04 from the preceding step (2.7 g, 8.16 mmoL) wasdissolved in dioxane (20 mL) and treated with freshly prepared 1N LiOH(9 mL). The reaction mixture was stirred at ambient temperature under N₂for 20 h. The reaction mixture was taken in EtOAc and washed with H₂O.The combined aqueous phase was cooled to 0° C. and acidified to pH 1.65using 1N HCl. The turbid mixture was extracted with EtOAc (2×100 mL).Combined organic layer was washed with brine, dried over MgSO₄, andconcentrated to give the desired acid, 23.05 (3.40 g).

Step 4:

To a suspension of NaBH(OAc)₃ (3.93 g, 18.5 mmoL) in CH₂Cl₂ (55 mL) wasadded a solution of product 23.05 from preceding step in anhydrousCH₂Cl₂ (20 mL) and acetic acid (2 mL). The slurry was stirred at ambienttemperature for 20 h. Ice cold water (100 mL) was added to the slurryand stirred for ½ hr. Organic layer was separated, filtered, dried andevaporated to yield the desired product, 23.06.

Step 5:

To a solution of the product 23.06 from preceding step (1.9 g) in MeOH(40 mL) was treated with excess of CH₂N₂/Et₂O solution and stirred forovernight. The reaction mixture was concentrated to dryness to yield acrude residue. The residue was chromatographed on silica gel, elutingwith a gradient of EtOAc/hexane to afford 1.07 g of the pure desiredproduct, 23.07.

Step 6:

To a solution of product 23.07 from preceding step (1.36 g) in anhydrousCH₂Cl₂ (40 mL) was treated with BF₃. Me₂O (0.7 mL). The reaction mixturewas stirred at ambient temperature for 20 h and quenched with sat.NaHCO₃ (30 mL) ad stirred for ½ hr. Organic layer was separated andcombined organic layer was washed with brine, dried over MgSO₄,concentrated to give crude residue. The residue was chromatographed onsilica gel eluting with a gradient of EtOAc/hexane to afford 0.88 g ofthe desired compound, 23.08.

Step 7:

To a solution of the product 23.08 (0.92 g) from preceding step in MeOH(30 mL) was added 10% Pd/C (0.16 g) at room temperature and hydrogenatedat ambient temperature under 1 atm. Pressure. The reaction mixture wasstirred for 4 h and concentrated to dryness to yield the desiredcompound, 23.01.

Preparation of P3 Moieties

Preparation of Intermediate 50.01

Step 1

To a solution of 5.02 (15 g) in MeOH (150 mL) was added conc HCl (3-4mL) and the mixture was refluxed for 16 h. The reaction mixture wascooled to room temperature and concentrated. The residue was taken indiethyl ether (250 mL) and washed with cold saturated sodium bicarbonatesolution, and brine. The organic layer was dried (Na₂SO₄) andconcentrated to afford the methyl ester 50.03 (12.98 g) which wascarried forward without further purification.

Step 2

The methyl ester 50.03 from above was dissolved in methylene chloride(100 mL) and cooled to −78° C., under nitrogen atmosphere. DIBAL (1.0 Msolution in methylene chloride, 200 mL) was added dropwise over 2 hperiod. The reaction mixture was warmed to room temperature over 16 h.The reaction mixture was cooled to 0° C. and MeOH (5-8 mL) was addeddropwise. A solution of aqueous 10% sodium potassium tartarate (200 mL)was slowly added with stirring. Diluted with methylene chloride (100 mL)and separated the organic layer (along with some white precipitate). Theorganic layer was washed with 1 N HCl (250 mL), brine (200 mL), dried(Na₂SO₄) and concentrated to provide the alcohol 50.04 (11.00 g) as aclear oil.

Step 3

The alcohol 50.04 from above was dissolved in methylene chloride (400mL) and cooled to 0° C. under nitrogen atmosphere. PCC (22.2 g) wasadded in portions and the reaction mixture was slowly warmed to roomtemperature over 16 h. The reaction mixture was diluted with diethylether (500 mL) and filtered through a pad of celite. The filtrate wasconcentrated and the residue was taken in diethyl ether (500 mL). Thiswas passed through a pad of silica gel and the filtrate was concentratedto provide the aldehyde 50.05 which was carried forward without furtherpurification.

Step 4

The aldehyde 50.05 from above was converted to the desired material50.01 using essentially the method of Chakraborty et. al (Tetrahedron,1995, 51(33), 9179-90).

PREPARATION OF SPECIFIC EXAMPLES Example 1 Synthesis of Compound ofFormula 10001

The amine, 10001a, (C. A. Busacca et al, Tetrahedron: Asymmetry; (2000)9 1907) (1.5 g, 6.9 mmol, 1 equiv.) was dissolved in dry dichloromethane20 ml) and cooled to −78° C. Added 3 ml (3 equiv.) of Et₃N followed bythe slow addition of dimethylsulfamyl chloride (1.5 eq., Sigma-Aldrich)dissolved in DCM. The temperature was kept at −78° C. until the additionis complete and then stirred overnight allowing it to raise to roomtemperature. Diluted with methylene chloride and washed with water, aq.1N HCl and finally brine. The organic layers were dried over anhydroussodium sulfate, filtered and concentrated in vacuo. Crude productisolated was purified via flash column (10→30% EtOAc-Hexane) to afford1.27 g (58%) of 10001b.

¹H NMR (CDCl₃, 300 MHz) δ, 4.6 (d, 1H), 3.45 (m, 1H), 3.25 (d, 1H), 2.89(s, 6H), 1.89 (bs, NH), 1.22 (s, 9H), 0.98 (s, 9H).

MS (ESI), m/z, relative intensity 324 [(M+1) 85], 268 (100), 224 (50).

Step B:

To the Boc protected sulfonyl urea 10001b (440 mg, 1.25 mmol, 1 equiv.)in DMF (10 mL) at 0° C. was added Cs₂CO₃ (613 mg, 1.5 equiv, 1.88 mmol)and MeI (6.36 mmol, 5 equiv., 0.601 mL) under inert atmosphere. Thereaction mixture was stirred at room temperature for 90 min and quenchedwith water. The aqueous layers were extracted with EtOAc, washed 4 timeswith water and brine. The organic layers was dried over anhydrous sodiumsulfate, filtered and evaporated off the solvent to afford 420 mg (91%)of 10001c that was used in the next reaction without furtherpurification.

¹H NMR (CDCl₃, 300 MHz) δ, 4.59 (d, 1H), 3.62-3.58 (m, 1H), 3.29-3.22(m, 1H), 2.80 (s, 3H), 2.79 (s, 6H), 1.89 (bs, NH), 1.22 (s, 9H), 0.98(s, 9H).

MS (ESI), m/z, relative intensity 338 [(M+1) 60], 282 (100), 238 (90)

Step C:

To the Boc-protected sulfonyl urea 10001c (890 mg, 1 equiv.) was added4M solution of HCl in dioxane (25 mL) at room temp and stirred for 1 hr.After the disappearance of starting material (TLC), the reaction mixturewas concentrated and azeotroped with hexanes and ether. The residue wastriturated with ether and the solid separating out was filtered anddried in vacuum to afford a pale yellow solid (720 mg, ˜100%). It wasused in further reaction without purification.

Step D:

To the amine hydrochloride salt 10001d (720 mg, 2.63 mmol) indichloromethane (15 ml) was added 15 ml of aq. saturated NaHCO₃ andstirred vigorously at 0° C. for 5 min. A solution of phosgene (2 equiv.20% in toluene) was syringed out to the lower layer and restored thevigorous stirring immediately. Checked the TLC at times and after 2 hrs,it showed complete consumption of starting material. The methylenechloride layer was separated and the aqueous layer was extracted withdichloromethane (30 ml). The combined organic layers were dried overanhydrous sodium sulfate, filtered and concentrated using rotaryevaporator under reduced pressure at rt. to half the volume and thenflushed N₂ for 15 minutes. Diluted the solution to 130 mL withdichloromethane and used as 0.02 M solution in further reactions.

Step E:

To the amine hydrochloride salt, 10001f (synthesized by coupling ofintermediate 10.12 and 1.17 using HATU followed by Dess-Martin oxidationand Boc deprotection following method C or the procedure outlined forthe synthesis of 13001h) (130 mg, 0.261 mmol, 1 equiv.) indichloromethane (5 ml) was added DIPEA (6 equiv.) at 0° C. A solution ofisocyanate 10001e (1 equiv, 13 ml of 0.02M soln.) under N₂ atmosphereand stirred for 30 min at ice temperature and 90 min at roomtemperature. The reaction mixture was quenched with citric acid andextracted with EtOAc. The organic layer was washed with brine, driedover anhydrous sodium sulfate, filtered and concentrated in vacuo. Thecrude product was purified using flash column chromatography (SiO₂,10-40% acetone-hexane) to afford 110 mg (59%) of 10001 as a colorlesssolid.

MS (ESI), m/z, relative intensity 724[(M+1) 45], 377 (100).

Example 2 Synthesis of Compound of Formula 10002

To a solution of amine hydrochloride salt 10002a (500 mg, 1.00 mmol, 1equiv.) in dichloromethane (15 ml) was added a solution of aq. sat.NaHCO₃ (15 mL). The reaction mixture was stirred vigorously at icetemperature for 5 min. A solution of phosgene (2 equiv. 20% in toluene)was syringed out to the lower layer and restored the vigorous stirringimmediately. Checked the TLC at times and after 2 hrs it showed completeconsumption of starting material and then separated the layers. Washedthe water layer one more time with DCM (3 ml) and dried over sodiumsulfate. Filtered and evaporated at high vacuum to half the volume andthen purged N₂ for 15 minutes. Used 10002b as a stock solution of 0.02Mby diluting with 50 mL of dichloromethane.

Step B:

To the ammonium salt 10001d, (80 mg, 0.293 mmol, 1 equiv.) indichloromethane (10 ml) was added DIPEA (6 equiv.) at ice temperature.Added isocyanate 10002b (1 equiv, 14.6 ml of 0.02M solution) under N₂atm., and stirred for 30 min at ice temperature and 90 ml at roomtemperature. Quenched with citric acid and extracted with EtOAc andwashed with brine. Dried over anhydrous sodium sulfate and filtered andevaporated off the solvent. The crude product was purified using silicagel flash chromatography (10-40% acetone-hexane) to afford 120 mg (57%)of 10002 as a colorless solid.

MS (ESI), m/z, relative intensity 724 [(M+1) 100], 461 (45), 403 (80).

Example 3 Synthesis of Compound of Formula 10003

To the amine hydrochloride 10003a, prepared as described before, (500mg, 1.03 mmol, 1 equiv.) in DCM (15 ml) was added 15 ml of sat. NaHCO₃.Stirred vigorously at ice temperature for 5 min. Stopped stirring andphosgene (1.11 mL, 2 equiv., 20% in toluene) was syringed out to thelower layer and restored the vigorous stirring immediately. Checked theTLC at times and after 2 hrs it showed complete consumption of startingmaterial and then separated the layers. Washed the water layer one moretime with DCM (3 ml) and dried over sodium sulfate. Filtered andevaporated at high vacuum to half the volume and then purged N₂ for 15minutes. Used as a stock solution of 10003b (0.02M) by diluting with 50mL of dichloromethane.

To a solution of ammonium salt, 10001d, (20 mg, 0.073 mmol, 1 equiv.) inDCM (5 ml) was added DIPEA (6 equiv.) at ice temperature was addedisocyanate 10003b (1 equiv, 0.073 mmol, 3.66 mL of 0.02M soln) under N₂atm. and stirred for 30 min at ice temperature and 90 min at roomtemperature. Quenched with citric acid and extracted with EtOAc andwashed with brine. Dried over anhydrous sodium sulfate, filtered andevaporated off the solvent. The crude product was purified via flashsilica column (10-40% acetone-hexane) to afford 28 mg, 55% of 10003.

MS (ESI), m/z, relative intensity 724 [(M+1) 100], 461 (45), 403 (80).

Example 4 Synthesis of Compound of Formula 10004

To 0.95 mL (11.7 mmol) of sulfuryl chloride in 20 mL ether was addeddropwise 2.3 mL (23.4 mmol) of piperidine at −78° C. The reaction wasstirred at rt for 3 hrs. The insoluble solid was removed by filtrationand the filtrate was washed with 1N HCl, sat NaHCO₃ and brine. Theorganic layer was dried over MgSO₄, filtered and the filtrate wasconcentrated to dryness to give 1.00 g of 10004b. yield 46%.

Step B:

To 0.350 g (1.62 mmol) of 10001a in 10 mL CH₂Cl₂ was added 0.23 mL (1.62mmol) of Et₃N, then 0.446 g (2.42 mmol) of 10004b in 5 mL CH₂Cl₂ dropwise at rt. The reaction mixture was stirred at rt overnight. Thereaction mixture was diluted with EtOAc, washed with a solution of aq.NH₄Cl and brine. The organic layers was dried over MgSO₄, filtered,concentrated in vacuo and purified by silica gel chromatography with6→24% EtOAc in Hexane to yield 0.353 g of product. Yield 60%.

Step C:

To 15 mg (0.041 mmol) of 10004c in a flask was added 2 ml (8 mmol) of 4MHCl (in dioxane) and stirred at RT for 50 min. The reaction mixture wasconcentrated to dryness in vacuo to give 32 mg of 2004d. Yield 100%.

Note: The conversion of 10004d to 10004 was identical to step B. inpreparative example 3 Synthesis of compound of formula 10003.

Example 5 Synthesis of Compound of Formula 10005

To a solution of 0.275 g (0.76 mmol) of 10004c in DMF was added 0.369 g(1.13 mmol) of Cs₂CO₃ and 0.085 mL (1.37 mmol) of MeI at 0° C. Thereaction was stirred at rt. overnight. The reaction mixture was dilutedwith water and extracted with EtOAc. The organic layer was washed withbrine, dried, concentrated in vacuo and the residue was purified bychromatography (SiO₂ 8˜32% EtOAc in hexane) to give 0.256 g of product10005a. Yield 89%.

Step B:

To a solution of 0.291 g (0.77 mmol) of 10005a in flask was added 3 mL(12 mmol) of 4M HCl (in dioxane). and stirred at rt. for 50 min. Afterthe completion of reaction as indicated by TLC the reaction mixture wasconcentrated in vacuo to dryness to yield 0.241 g of 10005b.

Step C:

To 0.241 g (0.77 mol) of 10005c in CH₂Cl₂ at 0° C. was added 0.81 mL(1.54 mmol) of 1.9 M solution of phosgene in toluene and 10 mL ofsaturated aqueous NaHCO₃ solution. The reaction mixture was stirred atrt. for 1.0 h. The organic layer was separated and dried over (Na₂SO₄),filtered concentrated to half volume. It was further diluted with CH₂Cl₂to afford a 0.07 M solution.

Note: The conversion of 10005c to 10005 was identical to step B inpreparative example 3 Synthesis of compounds 10003.

Compounds indicated in the following Table 1 were synthesized usingsimilar reactions as shown in Examples 1-5. Range of Ki* indicated: A≦75nM; 75<B≦250 nM; C>250 nM.

TABLE 1 Entry Structure K_(i)* 10001

A 10002

A 10003

A 10004

A 10005

A 10006

B 10007

A 10008

A 10009

B 10010

A 10011

A 10012

A 10013

A 10014

A 10015

A 10016

A 10017

A 10018

B 10019

A 10020

A 10021

A 10022

A 10023

A 10024

A 10025

A 10026

A 10027

A 10028

A 10229

A 10030

A 10031

A 10032

A 10033

A 10034

A 10035

B 10036

A 10037

A 10038

A 10039

A 10040

A 10041

A 10042

A 10043

A 10044

A 10045

A 10046

A 10047

A 10048

A 10049

A 10050

A 10051

A 10052

A 10053

A 10054

A 10055

A 10056

A 10057

A 10058

A 10059

A 10060

A 10061

A 10062

A 10063

A 10064

A 10065

A 10066

A 10067

A 10068

A 10069

A 10070

A 10071

A 10072

A 10073

A 10074

A 10075

A 10076

A 10077

A 10078

A 10079

A 10080

A 10081

A 10082

A 10083

A 10084

A 10085

A 10086

A 10087

A 10088

A 10089

A 10090

A 10091

A 10092

A 10093

A 10094

A 10095

A 10096

A 10097

A 10098

A 10099

A 10100

A 10101

A

Example-6 Synthesis of Compound of Formula 11001

A solution of amide 11001a (Obtained by Cbz protection oftert-butylglycine-N-methyl amide obtained from commercial source:TCI-Japan) 18 g, 64.67 mmol) in toluene (200 mL) was treated withBH₃.DMS (2 M soln. in THF, 65 mL, 130 mmol) and heated at 80° C. for 3h. The reaction mixture was cooled to rt. and treated carefully with aq.NaOH (2 M solution) and extracted into CH₂Cl₂ (3×200 mL). The combinedorganic layers were extracted with aq. saturated NaHCO₃ (3×300 mL),brine (300 mL), dried (MgSO₄) and purified by chromatography (SiO₂,ammoniacal methanol (7M)/CH₂Cl₂ 1:20) to yield 11001 b as a colorlessoil.

Step B:

A solution of amine 11001b (900 mg, 3.40 mmol) in CH₂Cl₂ at 0° C. wastreated with NMM (511 mg, 5.10 mmol) and triflic anhydride (585 mg, 5.10mmol) and stirred at 0° C. for 12 h. The reaction mixture was dilutedwith CH₂Cl₂ (300 mL) and washed with excess aq. HCl (1M, 500 mL). Theorganic layer was dried (MgSO₄) filtered concentrated in vacuo andpurified by chromatography (SiO₂, Hex/EtOAc 1:9-1:1) to yieldtrifluoromethane sulfonamide 11001c.

Step C:

A solution of 11001c (1.28 g, 3.22 mmol) in methanol (30 mL) was treatedwith palladium hydroxide (200 mg, 10% wt/C) and hydrogenated at 60 psifor 3 h. The reaction mixture was filtered through a plug of Celite® andthe filtrate was concentrated in vacuo. The residue was directly used infurther reaction without purification. A solution of deprotected amine(200 mg, 0.763 mmol) in DMF (3 mL), CH₂Cl₂ (3 mL) was treated with4-nitrophenylcarbamate 1.16 (409 mg, 0.915 mmol), NMM (308 mg, 3.05mmol) at 0° C. and stirred at rt. overnight. The reaction mixture wasconcentrated in vacuo diluted with CH₂Cl₂ (150 mL) and washed with aq.HCl (1M, 2×125 mL), aq. saturated NaHCO₃ (2×125 mL), brine (100 mL),dried (MgSO₄), filtered and purified by chromatography (SiO₂,CH₂Cl₂/EtOAc 1:19) to yield 11001d.

Step D:

A solution of methyl ester 11001d (210 mg, 0.368 mmol) in dry THF (3 mL)was treated with H₂O (3 mL), methanol (3 mL) and treated with LiOHmonohydrate (41.9 mg, 1 mmol) and stirred for 3 h at rt. The reactionmixture was acidified to pH-2 and extracted into CH₂Cl₂ (100 mL). Theorganic layer was washed with H₂O (100 mL), brine (100 mL) dried (MgSO₄)filtered concentrated in vacuo to yield acid that was used as it is inthe next reaction.

A solution the acid (50 mg, 0.089 mmol) in dry CH₂Cl₂ (2 mL) and DMF (2mL) was cooled to 0° C. and treated with amine 10.11 (20 mg, 0.116 mmol)HATU (57.03 mg, 0.15 mmol) and NMM (40.4 mg, 0.40 mmol). The reactionwas stirred at 0° C. for 36 h and concentrated in vacuo. The residue wasdissolved in CH₂Cl₂ (100 mL) and washed with aq HCl (1 M, 2×100 mL), aq.saturated NaHCO₃ (2×100 mL) brine (100 mL), dried (MgSO₄) filtered,concentrated in vacuo to yield 11001e that was used in the next reactionwithout further purification.

Step E:

A solution of 11001e (50 mg, 0.075 mmol) in toluene (3 mL) and DMSO (3mL) was treated with EDCl (134 mg, 0.703 mmol), and dichloroacetic acid(45.3 mg, 0.351 mmol, 30 μL) and stirred at rt. for 3 h. The reactionmixture was diluted with CH₂Cl₂ (60 mL) and washed with aq. saturatedNaHCO₃ (30 mL), aq. HCl (1 M, 30 mL), brine (30 mL), dried (MgSO₄)filtered, concentrated in vacuo and purified by chromatography (SiO₂,acetone/Hexanes 20-50% linear gradient) to yield 11001.

Example-7 Synthesis of Compound of Formula 11002

A solution of amine 11001b (4.0 g, 15.14 mmol) in CH₂Cl₂ (100 mL) wastreated with di-tert-butyldicarbonate (4.13 g, 18.91 mmol) and stirredat rt. for 12 h. The reaction mixture was concentrated in vacuo andpurified by chromatography (SiO₂, EtOAc/Hexanes 1:5) to yield Bocprotected amine.

A solution of the Boc protected compound in methanol was treated withpalladium hydroxide and hydrogenated at 60 psi for 12 h. The reactionmixture was filtered through a plug of Celite® and the filtrate wasconcentrated in vacuo. The residue 11002a was used in subsequent stepswithout further purification.

Step B:

A solution of amine 11002a (134 mg, 0.58 mmol) in acetonitrile (20 mL)was treated with 4-nitrophenylcarbamate 1.16 (260 mg, 0.58 mmol), NMM(177 mg, 1.74 mmol) at 0° C. and stirred at rt. overnight. The reactionmixture was concentrated in vacuo, diluted with CH₂Cl₂ (250 mL) andwashed with aq. HCl (1M, 2×125 mL), aq. saturated NaHCO₃ (2×125 mL),brine (100 mL), dried (MgSO₄) filtered and purified by chromatography(SiO₂, CH₂Cl₂/EtOAc 1:19) to yield 11002b (279 mg).

Step C:

A solution of 11002b (279 mg, 0.52 mmol) in 4 M HCl dioxane was stirredat rt for 2 h and concentrated in vacuo. The residue was used in furtherreaction as it is.

The ammonium salt (274 mg, 0.58 mmol) was dissolved in CH₂Cl₂:DMF (1:1)and cooled to 0° C. The reaction mixture was treated with 4 eq of Et₃N(233 mg, 2.33 mmol) and 2 eq of 2-pyridinesulfonyl chloride (248 mg, 1.2mmol) and stirred at rt overnight. The reaction mixture was washed withsaturated NaHCO₃, and the organic layer was extracted with CH₂Cl₂. Theorganic layer was dried with MgSO₄, filtered, and concentrated in vacuo.The crude product was purified using silica gel chromatography using aHorizon HPFC system (30%-->90% EtOAc/hexanes) to yield 240 mg of 11002c.

Step D:

11002c (240 mg, 0.41 mmol) was dissolved in THF and H₂O (3:1) andtreated with 2.5 eq of LiOH.H₂O. The reaction mixture was treated withMeOH until the solution turns homogeneous. The reaction mixture wasstirred at rt for approximately 3 hr. The reaction mixture was treatedwith 1 M aq HCl and concentrated in vacuo. The aqueous layer wasextracted with CH₂Cl₂, dried with MgSO₄, filtered, and concentrated invacuo. The crude was used in further couplings without any purification.

The acid (179 mg, 0.32 mmol) was dissolved in 1:1 CH₂Cl₂/DMF and cooledto 0° C. The reaction mixture was treated with 1.3 eq of deprotected11.01 (11.01 was deprotected by dissolving (200 mg, 0.61 mmol) in 10 mLof TFA and 3 mL of Me₂S and standing for 3 h. The reaction mixture wasconcentrated in vacuo and used as it is in further couplings) (238 mg,0.41 mmol) 3.5 eq of NMM (112 mg 1.1 mmol), and 1.5 eq of HATU (180 mg,0.47 mmol), and stored in the freezer (˜0° C.) overnight. The reactionmixture was concentrated under high vacuum, and the residue was treatedwith saturated NaHCO₃. The aqueous layer was extracted with CH₂Cl₂,dried with MgSO₄, filtered, and concentrated in vacuo.

Step E:

11002d (313 mg, 0.42 mmol) was dissolved in CH₂Cl₂ and treated with 3 eqof Dess-Martin periodinane (535 mg, 1.3 mmol). The reaction mixture wasstirred at rt for approximately 2 hr. The reaction mixture was dilutedwith 2:1 1 M NaHSO₃/saturated NaHCO₃, and the aqueous layer wasextracted with CH₂Cl₂. The organic layer was washed with 1 M NaHSO₃ andsaturated NaHCO₃, dried with MgSO₄, filtered, and concentrated in vacuo.The crude product was purified using silica gel chromatography with aHorizon HPFC system (20%-->60% acetone/hexanes) to yield 11002

Example-8 Synthesis of Compound of Formula 11003

A solution of ammonium salt 11002c (880 mg, 1.86 mmol) in dry methylenechloride was cooled to 0° C. and treated with triethylamine (0.5 mL,3.71 mmol) and 2-thiophenesulfonyl chloride (678 mg, 3.71 mmol) andstirred at 0° C. for 48 h. The reaction mixture was taken in methylenechloride and the organic layer was washed with aq. HCl (1 M soln.), andbrine. The combined organic layers were dried (MgSO₄) filteredconcentrated in vacuo and purified by chromatography (SiO₂,acetone/Hexanes 1:4) to yield 978 mg of 11003a as a colorless solid.

Step C:

A solution of 11003a (1.2 g, 2.22 mmol) in THF/H₂O was treated withLiOH.H₂O and stirred at rt. for 3 h. The reaction mixture was acidifiedwith 1 M aq. HCl and extracted with CH₂Cl₂. The combined organic layerswere dried (MgSO₄), filtered concentrated in vacuo and used as it is inthe next step.

A solution of acid (100 mg, 0.175 mmol) in dry CH₂Cl₂ (4 mL) and DMF (4mL) was cooled to 0° C. and treated with amine 12.04 (100 mg, 0.263mmol) HATU (100 mg, 0.263 mmol) and NMM (70.4 mg, 0.704 mmol). Thereaction was stirred at 0° C. for 14 h and concentrated in vacuo. Theresidue was dissolved in CH₂Cl₂ (100 mL) and washed with aq. HCl (1 M,2×100 mL), aq. saturated NaHCO₃ (2×100 mL) brine (100 mL), dried(MgSO₄), filtered, concentrated in vacuo to yield 11003b that was usedin the next reaction without further purification.

Step D:

A solution of alcohol 11003b (100 mg, 0.133 mmol) in dry CH₂Cl₂ (4 mL)was treated with Dess-Martin reagent (Dess, D. B.; Martin, J. C. J. Am.Chem. Soc. 1991, 113, 7277.) (150 mg, 0.345 mmol) and stirred at rt for2 h. The reaction mixture was diluted with aq. Na₂S₂O₃ (5%, 30 mL) andaq. saturated NaHCO₃ (30 mL) and stirred at rt. for 15 min. The reactionmixture was extracted with CH₂Cl₂ (100 mL) and the combined organiclayers were dried (MgSO₄), filtered, concentrated in vacuo and purifiedby chromatography (SiO₂, acetone/Hexanes 20%-55% linear gradient) toyield 11003.

Example-9 Synthesis of Compound of Formula 11004

A solution of amine 11001a (150 mg, 0.567 mmol) in CH₂Cl₂ (5 mL) wascooled to 0° C. and treated with NMM (100 mg, 100 mL). The reaction wastreated with tert-butyl sulfenylchloride (Sun, P; Weinreb, S. M.; Shang,M. J. Org. Chem. 1997, 62, 8604) (0.5 mL, 1.3 M soln in CH₂Cl₂) andstirred at rt. overnight. The reaction mixture was diluted with aq. HCl(1M, 30 mL) and extracted with CH₂Cl₂ (3×30 mL). The combined organiclayer was extracted with brine (30 mL) dried (MgSO₄) filteredconcentrated in vacuo and purified by chromatography (SiO₂,acetone/Hexanes) to yield 11004a.

Step B:

A solution of sulfenamide 11004a (2.00 g, 5.43 mmol) in CH₂Cl₂ (60 mL)was treated with MCPBA (2.34 g, 8.145 mmol, 60%) and stirred at rt. for1 h. The reaction mixture was diluted with aq. Na₂S₂O₃ (10%, 50 mL) andaq NaHCO₃ (saturated, 100 mL) and stirred at rt. for 30 min. Thereaction mixture was extracted with CH₂Cl₂ (150 mL) and the combinedorganic layers were washed with water, brine, dried (MgSO₄) filteredconcentrated in vacuo and purified by chromatography (SiO₂, EtOAc/Hex1:9→1:1) to yield 11004b.

Step C:

A solution of Cbz-protected compound 11004b (1.5 g, 3.90 mmol) inmethanol (25 mL) was treated with palladium hydroxide (10% on C) andhydrogenated at 60 psi for 1 h. The reaction mixture was filteredthrough a plug of celite and concentrated in vacuo. It was used forfurther reaction without any purification.

A solution of deprotected amine (1.00 g, 4.00 mmol) in acetonitrile (20mL) was treated with 4-nitrophenylcarbamate 1.16 (1.879 g, 4.20 mmol),NMM (1.062 g, 10.5 mmol) and stirred at rt. overnight. The reactionmixture was concentrated in vacuo diluted with CH₂Cl₂ (200 mL) andwashed with aq. HCl (1M, 2×125 mL), aq. saturated NaHCO₃ (2×125 mL),brine (100 mL), dried (MgSO₄), filtered, and purified by chromatography(SiO₂, CH₂Cl₂/EtOAc 1:19) to yield 11004c.

Step D:

Intermediate 11004c was converted to 11004 by coupling to intermediate12.03 followed by Moffett oxidation identical to the proceduresdescribed in preparative example-6 of synthesis of 11001, Step D andStep E. Compounds shown in the following Table 2 were synthesized usingsimilar reactions as shown in Examples above. Range of Ki* IndicatedA≦75 nM; 75<B≦250 nM; C>250 nM.

TABLE 2 Entry Structure Ki* 11001

A 11002

A 11003

A 11004

A 11005

B 11006

A 11007

A 11008

A 11009

C 11010

A 11011

A 11012

A 11013

A 11014

C 11015

A 11016

C 11017

B 11018

A 11019

A 11020

A 11021

A 11022

B 11023

B 11024

B 11025

A 11026

A 11027

A 11028

A 11029

A 11030

A 11031

A 11032

A 11033

A 11034

A 11035

A 11036

A 11037

B 11038

A 11039

A 11040

B 11041

A 11042

A 11043

A 11044

A 11045

A 11046

A 11047

A 11048

A 11049

A 11050

A 11051

11052

A 11053

A 11054

A 11055

A 11057

A 11058

A 11059

A 11060

A 11061

A 11062

A 11063

A 11064

A 11065

A 11066

A 11067

A 11068

A 11069

A 11070

A 11071

A 11072

A 11073

A 11074

A 11075

A 11076

A 11077

A 11078

A 11079

A 11080

A 11081

A 11082

A 11083

A 11084

C 11085

A 11086

B 11087

B 11088

A 11089

A 11090

A 11091

A 11092

A 11093

C 11094

A 11095

B 11096

B 11097

A 11098

B 11099

B 11100

A 11101

A 11102

A 11103

A 11104

A 11105

A 11106

A 11107

B 11108

A 11109

A 11110

A 11111

A 11112

A 11113

B 11114

A 11115

A 11116

A 11117

A 11118

A 11119

A 11120

A 11121

A 11122

A 11123

A 11124

A 11125

A 11126

A 11127

A 11128

A 11129

A 11130

A 11131

A 11132

B 11133

A 11134

A 11135

A 11136

B 11137

A 11138

A 11139

A 11140

A 11141

A 11142

A 11143

A 11144

A 11145

A 11146

A 11147

A 11148

A 11149

A 11150

A 11151

A 11152

A 11153

A 11154

A 11155

A 11156

A 11157

A 11158

A 11159

A 11160

A 11161

A 11162

A 11163

A 11164

A 11165

A 11166

A 11167

A 11168

A 11169

A 11170

A 11171

A 11172

A 11173

A 11174

A 11175

A 11176

A 11177

A 11178

A 11179

A 11180

A 11181

A 11182

A 11183

A 11184

A 11185

A 11186

A 11187

A 11188

A 11189

A 11190

A 11191

A 11192

A 11193

B 11194

B 11195

B 11196

A 11197

A 11198

A 11199

A 11200

A 11201

A 11202

A 11203

A 11204

B 11205

A 11206

B 11207

A 11208

A 11209

A 11210

A 11211

A 11212

A 11213

B 11214

A 11215

B 11216

B 11217

A 11218

A 11219

A 11220

A 11221

A 11222

A 11223

A 11224

B 11225

A 11226

A 11227

A 11228

A 11229

A 11230

A 11231

A 11232

A 11233

A 11234

A 11235

A 11236

A 11237

A 11238

A 11239

A 11240

A 11242

A 11243

B 11244

A 11245

A 11246

A 11247

A 11248

A 11249

A 11250

A 11251

A 11252

A 11253

A 11254

A 11255

A 11256

A 11257

A 11258

A 11259

A 11260

A 11261

A 11262

B 11263

A 11264

A 11265

A 11266

A 11267

A 11268

A 11269

A 11270

A 11271

A 11272

A 11273

B 11274

A 11275

A 11276

A 11277

A 11278

A 11279

A 11280

A 11281

A 11282

B 11283

C 11284

B 11285

A 11286

A 11287

A 11288

A 11289

A 11290

A 11291

A 11292

A 11293

A 11294

A 11295

A 11296

A 11297

A 11298

A 11299

A 11300

C 11311

C 11312

C 11313

C 11314

C 11315

A 11316

A 11317

A 11318

A 11319

A 11320

A 11321

A 11322

A 11323

A 11324

A 11325

A 11326

A 11327

A 11328

A 11329

A 11330

A 11331

A 11332

B 11333

B 11334

A 11335

A 11336

A 11337

A 11338

A 11339

A 11340

A 11341

A 11342

A 11343

A 11344

A 11345

A 11346

A 11347

A 11348

A 11349

A 11350

A 11351

A 11352

A 11353

A 11354

A 11355

A 11356

A 11357

A 11358

A 11359

A 11360

A 11361

A 11362

A 11363

A 11364

B 11365

A 11366

A 11367

A 11368

A 11369

B 11370

A 11371

A 11372

A 11373

A 11374

A 11375

A 11376

A 11377

A 11378

B 11379

A 11380

A 11381

A 11382

A 11383

A 11384

A 11385

A 11386

A 11387

A 11388

A 11389

C 11390

A 11391

B

Example 10 Synthesis of Compound of Formula 12001

-   -   12001a 12001b

A solution of 12001a (2.0 g, 9.2 mmol, Indofine chemicals) in toluene(150 mL) was treated with BH₃.DMS (˜10 M, 3 mL) and heated at 90° C. for2 h. The reaction mixture was cooled to 0° C. and diluted with 2 M aq.NaOH. The organic layer was extracted with CH₂Cl₂ and the combinedorganic layers were dried (MgSO₄) filtered concentrated in vacuo toyield 1.1 g of 12001 b.

Step B:

A solution of amine 12001b (500 mg, 2.5 mmol) in CH₂Cl₂ (10 mL) wastreated with benzene sulfonyl chloride (669 mg, 3.8 mmol) and Et₃N (384mg, 3.8 mmol) and stirred overnight at 0° C. The reaction mixture wasdiluted with aq. 1 M HCl and extracted with CH₂Cl₂. The combined organiclayer was dried, filtered and concentrated in vacuo. The residue waspurified by chromatography (SiO₂, EtOAc/Hexanes 1:3) to yield 552 mg ofboc protected phenyl sulfonamide compound.

A solution of Boc-compound (552 mg, 1.6 mmol) in 4M HCl in dioxane atrt. was stirred for 1 h and concentrated in vacuo. The residue wastriturated with ether and the solid separating out was isolated bydecanting the ether layer and dried in vacuum to yield 12001c.

Step C:

A solution of deprotected amine 12001c (139 mg, 0.5 mmol) in CH₂Cl₂/DMF(1:1, 20 mL) was treated with 4-nitrophenylcarbamate 1.16 (1.879 g, 4.20mmol), NMM (1.062 g, 10.5 mmol) and stirred at rt. overnight. Thereaction mixture was concentrated in vacuo diluted with CH₂Cl₂ (200 mL)and washed with aq. HCl (1M, 2×125 mL), aq. saturated NaHCO₃ (2×125 mL),brine (100 mL), dried (MgSO₄), filtered, and purified by chromatography(SiO₂, Hexanes/EtOAc 1:2) to yield 12001d.

Step D:

Intermediate 12001d was converted to 12001 by coupling to intermediate10.11 followed by Moffett oxidation identical to the proceduresdescribed in preparative example-6 of synthesis of 11001, Step D andStep E.

Example 11 Synthesis of Compound of Formula 12002

A solution 12002a (Gregory, H. et al.; J. Chem. Soc. 1968; 531) (11.62g, 42.08 mmol) in dry toluene was treated with BH₃.DMS (˜10 M soln, 6.3mL) and heated at 70° C. overnight. The reaction mixture was cooled tort and quenched with aq. NaOH and extracted with CH₂Cl₂. The combinedorganic layers were extracted with brine and concentrated in vacuo toyield 12002b 8.77 g (80%).

Step B:

A solution of 12002b (2 g, 7.24 mmol) in methylene chloride was treatedwith pyridine (7.9 g, 100 mmol) and methanesulfonyl chloride (1.24 g,10.86 mmol) and stirred at rt. for 24 h. The reaction mixture was washedwith aq HCl, saturated NaHCO₃ and brine. The organic layer was dried(MgSO₄) filtered, concentrated in vacuo and purified by chromatography(SiO₂, acetone/Hexanes 1:2) to yield 12002c.

Step C:

A solution of 12002c (465 mg, 1.37 mmol) in methanol was treated withpalladium on carbon and hydrogenated for 2 h at 50 psi. The reactionmixture was filtered through a plug of Celite® and concentrated in vacuoto isolate the deprotected amine which was used in the next reactionwithout further purification.

A solution of deprotected amine in CH₂Cl₂/DMF (1:1) was treated with4-nitrophenylcarbamate 1.16 (612 mg, 1.37 mmol), NMM (548 mg, 5.48 mmol)and stirred at rt. for 12 h. The reaction mixture was concentrated invacuo diluted with CH₂Cl₂ (200 mL) and washed with aq. HCl (1M, 2×125mL), aq. saturated NaHCO₃ (2×125 mL), brine (100 mL), dried (MgSO₄),filtered, and purified by chromatography (SiO₂, Hexanes/acetone 1:4) toyield 12002d (560 mg).

Step D:

Intermediate 12002d was converted to 12002 by coupling to intermediate12.04 followed by Dess-Martin oxidation identical to the proceduresdescribed in preparative example of synthesis of 11003, Step C and StepD.

Compounds shown in the following Table 3 were synthesized using similarreactions as shown in Examples above. Range of Ki* indicated: A≦75 nM;75<B≦250 nM; C>250 nM.

TABLE 3 Entry Structure Ki* 12001

A 12002

A 12003

A 12005

B 12006

B 12007

A 12008

A 12009

A 12010

A 12012

A 12013

A 12014

C 12015

C 12016

B 12017

A 12018

A 12019

A 12020

A 12021

A 12022

C 12023

A 12024

B 12025

C

Example-12 Synthesis of Compound of Molecular Formula 13001

The acid, 13001a, (5 g, 21.6 mmol, 1 equiv. Fluka) and methyl aminehydrochloride (1.2 equiv., 25.92 mmol) were dissolved in dryN,N-dimethyl formamide (20 ml) and cooled to 0° C. Added HATU (1.2equiv., 25.92 mmol) followed by DIPEA (Sigma-Aldrich), (172.8 mmol., 8equiv.) under N₂ atmosphere. The temperature was slowly raised to roomtemperature and stirred further for 4 h at room temperature. Dilutedwith EtOAc and washed with 1N HCl, NaHCO₃ and finally with brine. Driedover anhydrous sodium sulfate, filtered, and evaporated off the solvent.Crude product isolated was purified via flash column (10-50%EtOAc-Hexane) to afford 5.27 g of 13001b. Yield, (99%).

¹H NMR (CDCl₃, 300 MHz) δ, 6.0 (bs, 1H), 5.35 (d, 1H), 3.82 (d, 1H), 2.8(s, 3H), 1.4 (s, 9H), 0.98 (s, 9H).

Step B:

To the amide 13001b (3.37 g, 13.8 mmol, 1 equiv.) in toluene (100 mL) atroom temperature, added BH₃.Me₂S (10M, 3 equiv., 41.4 mmol, 4.14 mL) andrefluxed at 80° C. for 3 hrs. Evaporated off the solvent and the crudeproduct was quenched with 2 M aq. sodium hydroxide and extracted withdichloromethane. Washed the organic layer with brine and dried overanhydrous sodium sulfate. Filtered and evaporated off the solvent toafford 1.8 g of 13001c. The crude product was used for next step withoutpurification. Yield, (55%).

Step C:

To the Boc-protected amino compound 13001c (540 mg, 1 equiv.) indichloromethane (25 mL) at ice temp was added triethylamine (3 equiv.)and acetyl chloride (3 equiv.). Stirred for 1 hr at ice temperature andthen at room temperature for overnight Quenched with aq. sodiumbicarbonate and extracted with EtOAc. Washed with 1N HCl and then withbrine. The organic layer were dried over anhydrous sodium sulfate,filtered and evaporated off the solvent. The crude product was purifiedvia flash column (20-40% EtOAc-Hexane). Yield=230 mg (38%).

Step D:

To the amide 13001d (28 mg, 1 equiv.) added 4M HCl/dioxane (2 mL) atroom temperature. Stirred for 1 hr. TLC showed no starting material.Evaporated off the solvent and azeotroped with hexane and then withether. Washed out the non polar material with ether and kept under highvac. over the week end. Used the salt without purification; Productisolated (pale yellow solid)=22 mg (100%).

Step E:

To a mixture of acid, 1.17 (860 mg, 2.33 mmol, 1 equiv.) and aminehydrochloride 13.01 (570 mg, 2.56 mmol, 1.1 equiv.) in DMF (15 mL) atice temperature was added HATU (1.2 equiv., 1.066 g, 2.796 mmol) andDIPEA (8 equiv., 18.69 mmol, 3.26 mL) under N₂ and stirred at 0° C.overnight. The temperature was slowly allowed to rise to roomtemperature. Quenched with 1N HCl and extracted with EtOAc. The combinedorganic layer were washed with aq. NaHCO₃ (sat) and then with brine.Washed with ice cold water (5×20 ml) and again with brine. Dried overanhydrous Na₂SO₄, filtered, and evaporated off the solvent to afford1.25 g of 13001f. Yield, 100%.

Step F:

To the crude hydroxy amide, 13001f (2.71 mmol, 1.45 mg, 1 equiv.) in DCM(50 mL) at room temperature, was added Dess-Martin periodinane (2.30 mg,5.42 mmol, 2 equiv.) and stirred at rt. for 5 hrs. TLC showed completeconsumption of starting material and the appearance of product. Quenchedwith sat. NaHCO₃ aq thiosulfate solution and extracted with EtOAc. Theorganic layer was washed with brine and dried over anhydrous sodiumsulfate. Filtered and evaporated off the solvent. Crude product waspurified by silica gel flash column (10-40% acetone-hexane) to afford860 mg of 13001g; Yield, 62%.

Step G:

To the Boc amino compound 13001g (860 mg, 1 equiv.) added 4M HCl indioxane (25 mL) at room temperature. Stirred for 1 hr. TLC showed nostarting material. Evaporated off the solvent and azeotroped with hexaneand then with ether. Washed out the non polar material with ether andkept under high vacuum over the week end. Used the salt without furtherpurification; Product isolated (pale yellow solid)=750 mg (99%).

Step H:

To the ammonium salt, 13001 h, (150 mg, 0.318 mmol, 1 equiv.) in DCM (5ml) was added 5 ml of aq. sat. NaHCO₃. Stirred vigorously at icetemperature for 5 min. Stopped stirring and phosgene (2 equiv. 20% intoluene, 0.318 mL) was syringed out to the lower layer and restored thevigorous stirring immediately. Checked the TLC at times and after 2 hrsit showed complete consumption of starting material and then separatedthe layers. Washed the water layer one more time with DCM (3 ml) and thecombined organic layers were dried over sodium sulfate. The organiclayer was filtered and concentrated in vacuo to half the volume. Used13001i as a stock solution of 0.01M by diluting to 30 mL ofdichloromethane.

Step F:

To the ammonium salt 13001e (22 mg, 0.102 mmol, 1.1 equiv.) in DCM (10ml) was added DIPEA (6 equiv., 135 μL) at ice temperature. Addedisocyanate 13001i (1 equiv, 9 ml of 0.01M soln) under N₂ atm. andstirred for 30 min at ice temperature and 90 min at room temperature.Quenched with citric acid and extracted with EtOAc and washed withbrine. Dried over anhydrous sodium sulfate, filtered and evaporated offthe solvent. The crude product was purified via flash column (SiO₂,10-50% acetone-hexane) to afford 50 mg of 13001 as a colorless solid.Yield, (78%)

MS (ESI), m/z, 633 (M+1), 312.

Example 13 Synthesis of Compound of Molecular Formula 13002

To the Boc protected dipeptide 1.03 (3.6 g, 9.42 mmol, 1 equiv.) added4M HCl/dioxane (60 mL) at room temp. Stirred for 2 h. TLC showed nostarting material. Evaporated off the solvent and azeotroped with hexaneand then with ether. Washed out the non polar material with ether andkept under high vac. over night. Used the salt, 1.04, withoutpurification.

Product isolated=3 g (100%).

Step B:

To the amine hydrochloride 1.04 (3 g, 9.4 mmol) in dichloromethane (50ml) was added 50 ml of sat. NaHCO₃. Stirred vigorously at icetemperature for 5 min. Stopped stirring and phosgene (2 equiv. 20% intoluene, 10 mL) was syringed out to the lower layer and restored thevigorous stirring immediately. Checked the TLC at times and after 2 hrsit showed complete consumption of starting material and then separatedthe layers. Washed the water layer one more time with dichloromethane (3ml) and dried over anhydrous sodium sulfate. Filtered and evaporated offthe solvent using rotary evaporator under reduced pressure to half thevolume and then flushed N₂ for 15 minutes. Diluted to 33.5 mL withdichloromethane and used as 0.28 M solution for further couplings.

Step C

To the amine salt, 13001e, prepared as described before (151 mg, 0.73mmol, 1 equiv.) in DCM (10 ml) was added DIPEA (8 equiv., 1.01 mL, 5.84mmol) at ice temperature. Added isocyanate 13002a (1 equiv, 13 ml of0.02M soln) under N₂ atm. and stirred for 30 min at ice temperature and90 min at room temperature. Quenched with 10% citric acid and extractedwith EtOAc and washed with brine. Dried over anhydrous sodium sulfateand filtered and evaporated off the solvent. The crude product waspurified via flash column (20-80% EtOAC-hexane) to afford 270 mg of13002b as a white solid. Yield, (76%).

¹H NMR (CDCl₃, 300 MHz) δ, 5.8 (bs, NH), 5.4 (bs, NH), 5.2 (d, 1H), 4.4(d, 1H), 4-4.2 (m, 2H), 3.8-4 (m, 3H), 3.01 (s, 3H), 2.01 (bs, 6H), 1.6(m, 1H), 1.4 (m, 1H), 1.02-0.98 (m, 24H).

Step D

To the methyl ester, 13002b (270 mg, 0.562 mmol, 1 equiv.) in dioxane(10 ml) was added a solution of LiOH (10 equiv., 6 mL of 1N soln. inwater) and stirred overnight. Quenched with 1N HCl and extracted withEtOAC. Washed with brine and dried over anhydrous sodium sulfate.Filtered and evaporated off the solvent. crude yield 260 mg (99%).

Step E:

To the ammonium salt, 10.11 (16.06 mg, 0.077 mmol, 1.2 equiv.) in DCM(10 ml) was added 13002 c (30 mg, 0.064 mmol, 1 equiv.) and cooled to−20° C. and added HATU (1.2 equiv., 0.077 mmol, 29.37 mg) followed byDIPEA (8 equiv., 89.94 μL, 0.515 mmol). The reaction mixture was stirredovernight at that temperature. Quenched with 1N HCl and extracted withEtOAC. Washed the organic layer with aq. saturated sodium bicarbonateand then with brine. Dried over anhydrous sodium sulfate, filtered, andevaporated off the solvent. Purified via flash column (SiO₂, 10-90%EtOAc-Hexane) to afford 40 mg of hydroxyamide Yield, (100%).

To the hydroxyl amide (40 mg, 0.0645 mmol, 1 equiv.) in 1:1 mixture ofDMF/toluene (6 mL) at ice temperature was added EDCl.HCl (123 mg, 10equiv., 0.645 mmol) and dichloroacetic acid (27 μL, 5 equiv., 0.322mmol) and stirred for 5 min. and room temperature for 3 h. Quenched withbrine and washed with 1N HCl followed by aq. sat. NaHCO₃ and again withbrine. Dried over anhydrous sodium sulfate, filtered and evaporated offthe solvent. Purified via silica gel preparative TLC (50%acetone-hexane) to afford 30 mg of 13002. Yield (75%).

MS (ESI), m/z, 619 (M+1), 312.

Compounds shown in the following Table 4 were synthesized using similarreactions as shown in Examples above. Range of Ki* indicated A≦75 nM;75<B≦250 nM; C>250 nM.

TABLE 4 Entry Structure Ki* 13001

A 13002

A 13003

B 13004

A 13008

A 13009

A 13010

A 13011

A 13012

B 13013

A 13014

B 13015

B 13016

A 13017

C 13018

A 13019

A 13020

A 13021

A 13022

A 13023

A 13024

A 13025

A 13026

A 13027

A 13028

A 13029

A 13030

A 13031

A 13032

A 13033

A

Example 14 Synthesis of Compound of Formula 14001

A solution of 14001a (10.0 g, 40.0 mmol Indofine chemicals) in toluene(150 mL) was treated with BH₃.DMS (˜2 M, 40 mL) and heated at 90° C.overnight. The reaction mixture was cooled to 0° C. and diluted with 2 Maq. NaOH. The reaction mixture was heated at 90° C. for 15 min. Theaqueous layer was extracted with CH₂Cl₂ and the combined organic layerswere dried (MgSO₄) filtered concentrated in vacuo to yield 11 g of14001b.

Step B:

A solution of amine 14001b (10 g, 42.0 mmol) in CH₂Cl₂/DMF (1:5) wascooled to −78° C. and treated with di-tert-butyldicarbonate (13.8 g, 63mmol). The reaction mixture was stirred at rt for 48 h and diluted withaq. 1 M HCl and extracted with EtOAc. The combined organic layer waswashed with aq. NaHCO₃, brine, dried, filtered and concentrated invacuo. The residue was purified by chromatography (SiO₂, EtOAc/Hexanes1:3) to yield 4 g of Boc protected compound.

A solution of Boc-compound (6 g, 17.8 mmol) in methanol was treated withPd(OH)₂/C (1.89 g, 20% on C) and hydrogenated for 1 h. The reactionmixture was filtered through a plug of celite and concentrated in vacuo.The residue 14001c was used further reaction without purification.

Step C:

A solution of deprotected amine 14001c (3.6 g, 17.8 mmol) in CH₂Cl₂/DMF(1:1, 20 mL) was treated with 4-nitrophenylcarbamate 1.16 (7.97 g, 17.8mmol), NMM (4 g, 14.0 mmol) and stirred at rt. overnight. The reactionmixture was concentrated in vacuo diluted with EtOAc and washed with aq.HCl, aq. saturated NaHCO₃, brine, dried (MgSO₄), filtered, and purifiedby chromatography (SiO₂, Hexanes/EtOAc 3:7) to yield 14001 d

Step D:

A solution of 14001d was dissolved in 4 M HCl in dioxane and stirred atrt for 2 h. The reaction mixture was concentrated in vacuo and used infurther reactions without purification.

The ammonium salt (100 mg, 0.224 mmol) dissolved in DMF/CH₂Cl₂ (1:1) wastreated with isopropylchloroformate (54 mg, 0.448 mmol) and Et₃N (45 mg,0.448 mmol) at 0° C. and stirred at rt. overnight. The reaction mixturewas diluted with EtOAc and the organic layer was washed with aq. 1M HCl,aq. saturated NaHCO₃, and brine. It was dried (MgSO₄) filtered,concentrated in vacuo and used as it is in the next reaction.

Step E:

Intermediate 14001e was converted to 14001 by coupling to intermediate10.11 followed by Moffett oxidation identical to the proceduresdescribed in preparative example 6 of synthesis of 11001, Step D andStep E.

Example 15 Synthesis of Compound of Formula 14002

A solution of 14001d was dissolved in 4 M HCl in dioxane and stirred atrt for 2 h. The reaction mixture was concentrated in vacuo and used infurther reactions without purification.

The ammonium salt (100 mg, 0.224 mmol) dissolved in DMF/CH₂Cl₂ (1:1) wastreated with phenylisocyanate (53 mg, 0.448 mmol) and Et₃N (45 mg, 0.448mmol) at 0° C. and stirred at rt. overnight. The reaction mixture wasdiluted with EtOAc and the organic layer was washed with aq. 1M HCl, aq.saturated NaHCO₃, and brine. It was dried (MgSO₄) filtered, concentratedin vacuo and purified by chromatography (SiO₂, EtOAc/Hex 2:3) to yield14002a.

Step B:

Intermediate 14002a was converted to 14002 by coupling to intermediate10.11 followed by Moffett oxidation identical to the proceduresdescribed in preparative example 6 of synthesis of 11001, Step D andStep E.

Compounds shown in the following Table 5 were synthesized using similarreactions as shown in Examples above. Ki* Range Indicated A≦75 nM;75<B≦250 nM; C>250 nM.

TABLE 5 Entry Structure K_(i)* 14001

A 14002

A 14003

B 14004

A 14005

A 14009

A 14010

A 14011

A 14012

B 14016

B 14017

B 14019

A 14020

A 14021

A 14022

A 14023

B 14024

B 14025

C 14026

C 14027

B 14028

B 14029

C 14030

A 14031

B 14032

C 14033

B 14034

C 14035

A 14036

B 14037

C 14038

B 14039

C 14040

C 14041

C 14042

C 14043

C 14044

C 14045

C 14046

C 14047

C 14048

C 14049

C 14050

C 14051

C 14052

C 14053

C 14054

C 14055

C 14056

A 14057

A 14058

A 14059

A

Additional compounds of the present invention are shown in Table 5A:

TABLE 5A Entry Structure K_(i)* 11392

A 11393

A 11394

A 11395

A 11396

A 11397

B 11398

A 11399

A 11400

A 11401

A 11402

A 11403

A 11404

A 11405

A 11406

A 11407

A 11408

A 11409

A 11410

A 11411

A 11412

A 11413

A 11414

A 11415

A 11416

A 11417

A 11418

A 11419

A 11420

A 11421

A 11422

A 11423

A 11424

A 11425

A 11426

A Range of K_(i)* indicated A ≦ 75 nM; 75 < B ≦ 250 nM; C > 250 nM.

The present invention relates to novel HCV protease inhibitors. Thisutility can be manifested in their ability to inhibit the HCV NS2/NS4aserine protease. A general procedure for such demonstration isillustrated by the following in vitro assay.

Assay for HCV Protease Inhibitory Activity:

Spectrophotometric Assay: Spectrophotometric assay for the HCV serineprotease can be performed on the inventive compounds by following theprocedure described by R. Zhang et al, Analytical Biochemistry, 270(1999) 268-275, the disclosure of which is incorporated herein byreference. The assay based on the proteolysis of chromogenic estersubstrates is suitable for the continuous monitoring of HCV NS3 proteaseactivity. The substrates are derived from the P side of the NS5A-NS5Bjunction sequence (Ac-DTEDVVX(Nva), where X=A or P) whose C-terminalcarboxyl groups are esterified with one of four different chromophoricalcohols (3- or 4-nitrophenol, 7-hydroxy-4-methyl-coumarin, or4-phenylazophenol). Illustrated below are the synthesis,characterization and application of these novel spectrophotometric estersubstrates to high throughput screening and detailed kinetic evaluationof HCV NS3 protease inhibitors.

Materials and Methods:

Materials: Chemical reagents for assay related buffers are obtained fromSigma Chemical Company (St. Louis, Mo.). Reagents for peptide synthesiswere from Aldrich Chemicals, Novabiochem (San Diego, Calif.), AppliedBiosystems (Foster City, Calif.) and Perseptive Biosystems (Framingham,Mass.). Peptides are synthesized manually or on an automated ABI model431A synthesizer (from Applied Biosystems). UV/VIS Spectrometer modelLAMBDA 12 was from Perkin Elmer (Norwalk, Conn.) and 96-well UV plateswere obtained from Corning (Corning, N.Y.). The prewarming block can befrom USA Scientific (Ocala, Fla.) and the 96-well plate vortexer is fromLabline Instruments (Melrose Park, Ill.). A Spectramax Plus microtiterplate reader with monochrometer is obtained from Molecular Devices(Sunnyvale, Calif.).

Enzyme Preparation: Recombinant heterodimeric HCV NS3/NS4A protease(strain 1a) is prepared by using the procedures published previously (D.L. Sali et al, Biochemistry, 37 (1998) 3392-3401). Proteinconcentrations are determined by the Biorad dye method using recombinantHCV protease standards previously quantified by amino acid analysis.Prior to assay initiation, the enzyme storage buffer (50 mM sodiumphosphate pH 8.0, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside and10 mM DTT) is exchanged for the assay buffer (25 mM MOPS pH 6.5, 300 mMNaCl, 10% glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5 μM DTT)utilizing a Biorad Bio-Spin P-6 prepacked column.

Substrate Synthesis and Purification: The synthesis of the substrates isdone as reported by R. Zhang et al, (ibid.) and is initiated byanchoring Fmoc-Nva-OH to 2-chlorotrityl chloride resin using a standardprotocol (K. Barlos et al, Int. J. Pept. Protein Res., 37 (1991),513-520). The peptides are subsequently assembled, using Fmoc chemistry,either manually or on an automatic ABI model 431 peptide synthesizer.The N-acetylated and fully protected peptide fragments are cleaved fromthe resin either by 10% acetic acid (HOAc) and 10% trifluoroethanol(TFE) in dichloromethane (DCM) for 30 min, or by 2% trifluoroacetic acid(TFA) in DCM for 10 min. The combined filtrate and DCM wash isevaporated azeotropically (or repeatedly extracted by aqueous Na₂CO₃solution) to remove the acid used in cleavage. The DCM phase is driedover Na₂SO₄ and evaporated.

The ester substrates are assembled using standard acid-alcohol couplingprocedures (K. Holmber et al, Acta Chem. Scand., B33 (1979) 410-412).Peptide fragments are dissolved in anhydrous pyridine (30-60 mg/ml) towhich 10 molar equivalents of chromophore and a catalytic amount (0.1eq.) of para-toluenesulfonic acid (pTSA) were added.Dicyclohexylcarbodiimide (DCC, 3 eq.) is added to initiate the couplingreactions. Product formation is monitored by HPLC and can be found to becomplete following 12-72 hour reaction at room temperature. Pyridinesolvent is evaporated under vacuum and further removed by azeotropicevaporation with toluene. The peptide ester is deprotected with 95% TFAin DCM for two hours and extracted three times with anhydrous ethylether to remove excess chromophore. The deprotected substrate ispurified by reversed phase HPLC on a C3 or C8 column with a 30% to 60%acetonitrile gradient (using six column volumes). The overall yieldfollowing HPLC purification can be approximately 20-30%. The molecularmass can be confirmed by electrospray ionization mass spectroscopy. Thesubstrates are stored in dry powder form under desiccation.

Spectra of Substrates and Products: Spectra of substrates and thecorresponding chromophore products are obtained in the pH 6.5 assaybuffer. Extinction coefficients are determined at the optimal off-peakwavelength in 1-cm cuvettes (340 nm for 3-Np and HMC, 370 nm for PAP and400 nm for 4-Np) using multiple dilutions. The optimal off-peakwavelength is defined as that wavelength yielding the maximum fractionaldifference in absorbance between substrate and product (productOD−substrate OD)/substrate OD).

Protease Assay: HCV protease assays are performed at 30° C. using a 200μl reaction mix in a 96-well microtiter plate. Assay buffer conditions(25 mM MOPS pH 6.5, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, 5μM EDTA and 5 μM DTT) are optimized for the NS3/NS4A heterodimer (D. L.Sali et al, ibid.)). Typically, 150 μl mixtures of buffer, substrate andinhibitor are placed in wells (final concentration of DMSO≦4% v/v) andallowed to preincubate at 30° C. for approximately 3 minutes. Fifty μlsof prewarmed protease (12 nM, 30° C.) in assay buffer, is then used toinitiate the reaction (final volume 200 μl). The plates are monitoredover the length of the assay (60 minutes) for change in absorbance atthe appropriate wavelength (340 nm for 3-Np and HMC, 370 nm for PAP, and400 nm for 4-Np) using a Spectromax Plus microtiter plate readerequipped with a monochrometer (acceptable results can be obtained withplate readers that utilize cutoff filters). Proteolytic cleavage of theester linkage between the Nva and the chromophore is monitored at theappropriate wavelength against a no enzyme blank as a control fornon-enzymatic hydrolysis. The evaluation of substrate kinetic parametersis performed over a 30-fold substrate concentration range (˜6-200 μM).Initial velocities are determined using linear regression and kineticconstants are obtained by fitting the data to the Michaelis-Mentenequation using non-linear regression analysis (Mac Curve Fit 1.1, K.Raner). Turnover numbers (k_(cat)) are calculated assuming the enzyme isfully active.

Evaluation of Inhibitors and Inactivators: The inhibition constants(K_(i)) for the competitive inhibitors Ac-D-(D-Gla)-L-I-(Cha)-C—OH (27),Ac-DTEDVVA(Nva)-OH and Ac-DTEDVVP(Nva)-OH are determined experimentallyat fixed concentrations of enzyme and substrate by plotting v_(o)/v_(i)vs. inhibitor concentration ([I]_(o)) according to the rearrangedMichaelis-Menten equation for competitive inhibition kinetics:v_(o)/v_(i)=1+[I]_(o)/(K_(i) (1+[S]_(o)/K_(m))), where v_(o) is theuninhibited initial velocity, v_(i) is the initial velocity in thepresence of inhibitor at any given inhibitor concentration ([I]_(o)) and[S]_(o) is the substrate concentration used. The resulting data arefitted using linear regression and the resulting slope,1/(K_(i)(1+[S]_(o)/K_(m)), is used to calculate the K_(i) value. The KI*values for some of the inventive compounds are given in Table 6.

While the present invention has been described with in conjunction withthe specific embodiments set forth above, many alternatives,modifications and other variations thereof will be apparent to those ofordinary skill in the art. All such alternatives, modifications andvariations are intended to fall within the spirit and scope of thepresent invention.

1. A compound, or enantiomer, stereoisomer, rotamer, tautomer,diastereomer and racemate of said compound, or a pharmaceuticallyacceptable salt or ester of said compound, said compound of the generalstructure shown in Formula I:

wherein: R¹ is OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can bethe same or different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroalkyl-,cycloalkyl-, and arylalkyl-; or R¹⁰ is R¹⁴, wherein R¹⁴ is selected fromthe group consisting of:

A and M are connected to each other (in other words, A-E-L-M takentogether) such that the moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl; E is C(H)or C(R); L is C(H) or C(R); R, R²,and R³ can be the same or different, each being independently selectedfrom the group consisting of H, alkyl-, alkenyl-, alkynyl-, cycloalkyl-,heteroalkyl-, aryl-, (cycloalkyl)alkyl-, and aryl-alkyl-; Y is:

wherein Y³⁰ and Y³¹ are selected from

G is NH or O; and R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, T₁, T₂, T₃ and T₄ can be thesame or different, each being independently selected from the groupconsisting of H, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, aryl, and arylalkyl, or alternately, R¹⁷ andR¹⁸ are connected to each other to form a three to eight-memberedcycloalkyl; wherein each of said alkyl, aryl, or cycloalkyl or can beunsubstituted or optionally independently substituted with one or moremoieties selected from the group consisting of: hydroxy, alkoxy,aryloxy, thio, alkylthio arylthio, amino, amido, alkylamino, arylamino,alkylsulfonyl, arylsulfonyl, sulfonamido, alkyl, aryl, heteroaryl,alkylsulfonamido, arylsulfonamido, keto, carboxy, carbalkoxy,carboxamido, alkoxycarbonylamino, alkoxycarbonyloxy, alkylureido,arylureido, halo, cyano, and nitro.
 2. The compound of claim 1, whereinR¹ is NR⁹R¹⁰, and R⁹ is H, R¹⁰ is H, or R¹⁴ wherein R¹⁴ is selected fromthe group consisting of


3. The compound of claim 1, wherein R² is selected torn the groupconsisting of the following moieties:


4. The compound of claim 1, wherein R³ is selected from the groupconsisting of:

wherein R³¹ is OH or O-alkyl; and R³² is H, O(O)CH₃, C(O)OtBu orC(O)N(H)tBu.
 5. The compound of claim 4, wherein R³ is selected from thegroup consisting of the following moieties:


6. The compound of claim 1, wherein G is NH.
 7. The compound of claim 6,wherein Y is selected from the following moieties:

wherein Y³² is selected from the group consisting of:

Y³⁰ and Y³¹ is selected from

wherein u is a number 0-6; and R¹⁹ is selected from H, alkyl, phenyl orbenzyl.
 8. The compound of claim 7, wherein T₁, T₂ and T₃ can be thesame or different, each being independently selected from the groupconsisting of:


9. The compound of claim 1, wherein the moiety:

is selected from the following structures:


10. The compound of claim 9, wherein the moiety:

is selected from the following structures:


11. The compound of claim 10, wherein the moiety:

is selected from the following structures:


12. The compound of claim 1, wherein R¹ is NHR¹⁴, where R¹⁴ is selectedfrom the group consisting of:

R² is selected from the group consisting of the following moieties:

R³ is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

and the moiety:


13. A pharmaceutical composition comprising as an active ingredient ateast one compound of claim
 1. 14. The pharmaceutical composition ofclaim 13 for use in treating an infection of Hepatitis C Virus (“HCV”).15. The pharmaceutical composition of claim 14 additionally comprisingat least one pharmaceutically acceptable carrier.
 16. The pharmaceuticalcomposition of claim 15, additionally containing at least one antiviralagent.
 17. The pharmaceutical composition of claim 16, additionallycontaining at least one interferon.
 18. The pharmaceutical compositionof claim 17, wherein said at least one antiviral agent is ribavirin andsaid at least one interferon is α-interferon or pegylated interferon.19. A method of treating an infection of Hepatitis C Virus (“HCV”), saidmethod comprising administering to a patient in need of such treatment apharmaceutical composition which comprises therapeutically effectiveamounts of at least one compound of claim
 1. 20. The method of claim 19,wherein said administration is oral or subcutaneous.
 21. A method ofpreparing a pharmaceutical composition for treating an infection ofHepatitis C Virus (“HCV”), said method comprising bringing into intimatephysical contact at least one compound of claim 1 and at least onepharmaceutically acceptable carrier.
 22. A compound exhibiting HepatitisC Virus (“HCV”) protease inhibitory activity, or enantiomer,stereoisomer, rotamer, tautomer, diastereomer, and racemate of saidcompound, or a pharmaceutically acceptable salt or ester of saidcompound, said compound being selected from the compounds of structureslisted below:


23. A pharmaceutical composition for treating an infection of HepatitisC Virus (“HCV”), said composition comprising therapeutically effectiveamount of one or more compounds in claim 22 and a pharmaceuticallyacceptable carrier.
 24. The pharmaceutical composition of claim 23,additionally containing at least one antiviral agent.
 25. Thepharmaceutical composition of claim 24, additionally containing at leastone interferon or PEG-interferon alpha conjugate.
 26. The pharmaceuticalcomposition of claim 25, wherein said at least one antiviral agent isribavirin and said at least one interferon is α-interferon or pegylatedinterferon.
 27. A method of treatment of an infection of Hepatitis CVirus (“HCV”), comprising administering an effective amount of one ormore compounds of claim
 22. 28. A method of treating, or amelioratingone or more symptoms of hepatitis C, comprising administering atherapeutically effective amount of one or more compounds of claim 22.29. The method of claim 28, wherein the HCV protease is the NS3/NS4aprotease.
 30. The method of claim 29, wherein the compound or compoundsinhibit HCV NS3/NS4a protease.
 31. A method of treating an infection ofhepatitis C Virus (“HCV”), said method comprising administering to apatient in need of such treatment, a pharmaceutical composition whichcomprises therapeutically effective amounts of at least one compound, orenantiomers, stereoisomers, rotamers, tautomers, diastereomers, andracemates of said compound, or a pharmaceutically acceptable salt- orester of said compound, said compound being selected from the following:


32. A compound of claim 1 in purified form.
 33. The compound of claim 1,wherein Y is: